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1.
J Arthroplasty ; 2024 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-38677343

RESUMO

BACKGROUND: Periprosthetic joint infection (PJI) treatment has high failure rates even after 2-stage revision. Risk factors for treatment failure (TF) after staged revision for PJI are not well defined, nor is it well established how they correlate with the risks of developing an index PJI. Identifying modifiable risk factors may allow preoperative optimization, while identifying nonmodifiable risk factors can influence surgical options or advise against further surgery. We performed a systematic review and meta-analysis to better define predictors of TF in 2-stage revision for PJI. METHODS: The PubMed, Embase, and Scopus databases were searched from their inception in December 1976 to April 15, 2023. Studies comparing patient-related variables between patients successfully treated who had 2-staged revision total hip arthroplasty (THA) and patients with persistent infections were included. Studies were screened, and 2 independent reviewers extracted data, while a third resolved discrepancies. Meta-analysis was performed on these data. There were 10,052 unique studies screened, and 21 studies met the inclusion criteria for data extraction. RESULTS: There was good-quality evidence that obesity, liver cirrhosis, and previous failed revisions for PJI are nonmodifiable risk factors, while intravenous drug use (IVDU) and smoking are modifiable risk factors for TF after 2-stage revision for hip PJI. Reoperation between revision stages was also significantly associated with an increased risk of TF. Interestingly, other risk factors for an index PJI including male gender, American Society of Anesthesiology score, diabetes mellitus, and inflammatory arthropathy did not predict TF. Evidence on Charlson Comorbidity Index was limited. CONCLUSIONS: Patients with a smoking history, obesity, IVDU, previous failed revision for PJI, reoperation between stages, and liver cirrhosis are more likely to experience TF after 2-stage revision THA for PJI. Modifiable risk factors include smoking and IVDU and these patients should be referred to services for cessation as early as possible before 2-stage revision THA.

2.
Bone Joint J ; 106-B(5 Supple B): 74-81, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38688495

RESUMO

Aims: The aim of this study was to compare the biomechanical models of two frequently used techniques for reconstructing severe acetabular defects with pelvic discontinuity in revision total hip arthroplasty (THA) - the Trabecular Metal Acetabular Revision System (TMARS) and custom triflange acetabular components (CTACs) - using virtual modelling. Methods: Pre- and postoperative CT scans from ten patients who underwent revision with the TMARS for a Paprosky IIIB acetabular defect with pelvic discontinuity were retrospectively collated. Computer models of a CTAC implant were designed from the preoperative CT scans of these patients. Computer models of the TMARS reconstruction were segmented from postoperative CT scans using a semi-automated method. The amount of bone removed, the implant-bone apposition that was achieved, and the restoration of the centre of rotation of the hip were compared between all the actual TMARS and the virtual CTAC implants. Results: The median amount of bone removed for TMARS reconstructions was significantly greater than for CTAC implants (9.07 cm3 (interquartile range (IQR) 5.86 to 21.42) vs 1.16 cm3 (IQR 0.42 to 3.53) (p = 0.004). There was no significant difference between the median overall implant-bone apposition between TMARS reconstructions and CTAC implants (54.8 cm2 (IQR 28.2 to 82.3) vs 56.6 cm2 (IQR 40.6 to 69.7) (p = 0.683). However, there was significantly more implant-bone apposition within the residual acetabulum (45.2 cm2 (IQR 28.2 to 72.4) vs 25.5 cm2 (IQR 12.8 to 44.1) (p = 0.001) and conversely significantly less apposition with the outer cortex of the pelvis for TMARS implants compared with CTAC reconstructions (0 cm2 (IQR 0 to 13.1) vs 23.2 cm2 (IQR 16.4 to 30.6) (p = 0.009). The mean centre of rotation of the hip of TMARS reconstructions differed by a mean of 11.1 mm (3 to 28) compared with CTAC implants. Conclusion: In using TMARS, more bone is removed, thus achieving more implant-bone apposition within the residual acetabular bone. In CTAC implants, the amount of bone removed is minimal, while the implant-bone apposition is more evenly distributed between the residual acetabulum and the outer cortex of the pelvis. The differences suggest that these implants used to treat pelvic discontinuity might achieve short- and long-term stability through different biomechanical mechanisms.


Assuntos
Acetábulo , Artroplastia de Quadril , Prótese de Quadril , Desenho de Prótese , Reoperação , Tantálio , Tomografia Computadorizada por Raios X , Humanos , Acetábulo/cirurgia , Artroplastia de Quadril/métodos , Artroplastia de Quadril/instrumentação , Feminino , Masculino , Idoso , Estudos Retrospectivos , Pessoa de Meia-Idade , Fenômenos Biomecânicos , Idoso de 80 Anos ou mais , Falha de Prótese , Porosidade
3.
Knee ; 48: 105-119, 2024 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-38565037

RESUMO

BACKGROUND: Chondroprogenitors, with enhanced chondrogenic potential, have emerged to be a promising alternative for cell-based therapy in cartilage repair. Platelet-rich plasma (PRP), widely used for intra-articular treatment, has a short half-life. Freeze-dried PRP (FD-PRP), with an extended half-life and retained growth factors, is gaining attention. This study compares the efficacy of Migratory Chondroprogenitors (MCPs) in gelled PRP and FD-PRP using in-vitro and ex-vivo models, assessing FD-PRP as a potential off-the-shelf option for effective cartilage repair. METHODOLOGY: MCPs were isolated from osteoarthritic cartilage samples (n = 3), characterized through FACS and RT-PCR. For in-vitro analysis, cells were loaded into gelled PRP and FD-PRP scaffolds at a density of 1x106 cells per scaffold. Trilineage differentiation studies and live-dead assays were conducted on MCPs using Calcein AM/Propidium Homodimer-1. In ex-vivo analysis, MCPs of the same density were added to Osteochondral Units (OCU) with chondral defects containing PRP gel and FD-PRP scaffolds, harvested on the 15th and 35th days for histological examination. Controls included cell-free scaffolds. RESULTS: Our in-vitro analysis demonstrates the robust viability of MCPs in both scaffolds, with no discernible impact on their differentiation capacity. Ex-vivo analysis of the OCU for cartilage repair showed that the chondrogenic potential characterized by the accumulation of extracellular matrix containing glycosaminoglycans and collagen type II production (with no alteration in collagen type X), was observed to be better with the gel PRP and the gel PRP containing MCP groups. CONCLUSIONS: These findings support the preference for gel PRP as a superior synergistic scaffold for chondroprogenitor delivery.

4.
Arthroplast Today ; 23: 101218, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37841451

RESUMO

Hip prosthetic joint infection management is complex and expensive, especially in severe bone loss. Reducing the price of interval prosthesis when performing staged revision could minimize costs without compromising outcomes. We present 2 similar techniques developed independently that use an antibiotic-coated cephalomedullary nail with a total hip arthroplasty bearing (head and cemented acetabular component) attached to it as an interval proximal femoral replacement prosthesis. Using this technique, the femoral implant cost was reduced up to 10-fold. All patients have recovered well with resolution of infection and functional recovery similar to patients undergoing proximal femoral replacement. In one case, the lag screw (femoral neck) fractured at 5 months prompting the second-stage revision. This complication should be considered when deciding the timing of second-stage revisions in these cases.

5.
PLoS One ; 18(4): e0285106, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37104525

RESUMO

Obtaining regeneration-competent cells and generating high-quality neocartilage are still challenges in articular cartilage tissue engineering. Although chondroprogenitor cells are a resident subpopulation of native cartilage and possess a high capacity for proliferation and cartilage formation, their potential for regenerative medicine has not been adequately explored. Fetal cartilage, another potential source with greater cellularity and a higher cell-matrix ratio than adult tissue, has been evaluated for sourcing cells to treat articular disorders. This study aimed to compare cartilage resident cells, namely chondrocytes, fibronectin adhesion assay-derived chondroprogenitors (FAA-CPCs) and migratory chondroprogenitors (MCPs) isolated from fetal and adult cartilage, to evaluate differences in their biological properties and their potential for cartilage repair. Following informed consent, three human fetal and three adult osteoarthritic knee joints were used to harvest the cartilage samples, from which the three cell types a) chondrocytes, b) FAA-CPCs, and MCPs were isolated. Assessment parameters consisted of flow cytometry analysis for percentage expression of cell surface markers, population doubling time and cell cycle analyses, qRT-PCR for markers of chondrogenesis and hypertrophy, trilineage differentiation potential and biochemical analysis of differentiated chondrogenic pellets for total GAG/DNA content. Compared to their adult counterparts, fetal cartilage-derived cells displayed significantly lower CD106 and higher levels of CD146 expression, indicative of their superior chondrogenic capacity. Moreover, all fetal groups demonstrated significantly higher levels of GAG/DNA ratio with enhanced uptake of collagen type 2 and GAG stains on histology. It was also noted that fetal FAA CPCs had a greater proliferative ability with significantly higher levels of the primary transcription factor SOX-9. Fetal chondrocytes and chondroprogenitors displayed a superior propensity for chondrogenesis when compared to their adult counterparts. To understand their therapeutic potential and provide an important solution to long-standing challenges in cartilage tissue engineering, focused research into its regenerative properties using in-vivo models is warranted.


Assuntos
Cartilagem Articular , Condrócitos , Humanos , Adulto , Condrócitos/metabolismo , Condrogênese , Células Cultivadas , Cartilagem Articular/metabolismo , Diferenciação Celular , DNA/metabolismo
6.
J Orthop ; 35: 43-53, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36387762

RESUMO

Purpose of research: The potential for cartilage repair using articular cartilage derived chondroprogenitors has recently gained popularity due to promising results from in-vitro and in-vivo studies. Translation of results from in-vitro to a clinical setting requires a sufficient number of animal studies displaying significant positive outcomes. Thus, this systematic review comprehensively discusses the available literature (January 2000-March 2022) on animal models employing chondroprogenitors for cartilage regeneration, highlighting the results and limitations associated with their use.As per Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines, a web-based search of PubMed and SCOPUS databases was performed for the following terminologies: "chondroprogenitors", "cartilage-progenitors", and "chondrogenic-progenitors", which yielded 528 studies. A total of 12 studies met the standardized inclusion criteria, which included chondroprogenitors derived from hyaline cartilage isolated using fibronectin adhesion assay (FAA) or migratory assay from explant cultures, further analyzing the role of chondroprogenitors using in-vivo animal models. Principal results: Analysis revealed that FAA chondroprogenitors demonstrated the ability to attenuate osteoarthritis, repair chondral defects and form stable cartilage in animal models. They displayed better outcomes than bone marrow-derived mesenchymal stem cells but were comparable to chondrocytes. Migratory chondroprogenitors also demonstrated superiority to BM-MSCs in terms of higher chondrogenesis and lower hypertrophy, although a direct comparison to FAA-CPs and other cell types is warranted. Major conclusions: Chondroprogenitors exhibit superior properties for chondrogenic repair; however, limited data on animal studies necessitates further studies to optimize their use before clinical translation for neo-cartilage formation.

7.
JBJS Case Connect ; 12(4)2022 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-36820637

RESUMO

CASE: We report a case of acetabular reconstruction for a large defect with pelvic discontinuity that underwent 4 revisions for dislocations over a 3-year period. This allowed assessment of implant stability both on imaging, using measurements on plain radiographs and radiostereometric analysis (RSA) against both ilium and ischium, and direct assessment during each surgery. Only implant stability measured with RSA correlated with intraoperative revision findings. CONCLUSION: This case underlines the role of RSA in assessing early acetabular implant stability in pelvic discontinuity and the importance of assessing the stability of the implant against both ilium and ischium.


Assuntos
Artroplastia de Quadril , Prótese de Quadril , Humanos , Artroplastia de Quadril/métodos , Análise Radioestereométrica , Reoperação/métodos , Acetábulo/cirurgia
8.
Sci Rep ; 11(1): 23685, 2021 12 08.
Artigo em Inglês | MEDLINE | ID: mdl-34880351

RESUMO

Cell-based therapy for articular hyaline cartilage regeneration predominantly involves the use of mesenchymal stem cells and chondrocytes. However, the regenerated repair tissue is suboptimal due to the formation of mixed hyaline and fibrocartilage, resulting in inferior long-term functional outcomes. Current preclinical research points towards the potential use of cartilage-derived chondroprogenitors as a viable option for cartilage healing. Fibronectin adhesion assay-derived chondroprogenitors (FAA-CP) and migratory chondroprogenitors (MCP) exhibit features suitable for neocartilage formation but are isolated using distinct protocols. In order to assess superiority between the two cell groups, this study was the first attempt to compare human FAA-CPs with MCPs in normoxic and hypoxic culture conditions, investigating their growth characteristics, surface marker profile and trilineage potency. Their chondrogenic potential was assessed using mRNA expression for markers of chondrogenesis and hypertrophy, glycosaminoglycan content (GAG), and histological staining. MCPs displayed lower levels of hypertrophy markers (RUNX2 and COL1A1), with normoxia-MCP exhibiting significantly higher levels of chondrogenic markers (Aggrecan and COL2A1/COL1A1 ratio), thus showing superior potential towards cartilage repair. Upon chondrogenic induction, normoxia-MCPs also showed significantly higher levels of GAG/DNA with stronger staining. Focused research using MCPs is required as they can be suitable contenders for the generation of hyaline-like repair tissue.


Assuntos
Regeneração Óssea , Cartilagem Articular/fisiologia , Condrogênese , Fibronectinas/metabolismo , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Adipogenia , Biomarcadores , Ciclo Celular , Diferenciação Celular , Movimento Celular , Células Cultivadas , Condrócitos/citologia , Condrócitos/metabolismo , Imunofluorescência , Humanos , Pessoa de Meia-Idade
9.
Rev Bras Ortop (Sao Paulo) ; 56(5): 579-587, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34733429

RESUMO

Objective The management of tibia fractures complicated by compartment syndrome affects the treatment and functional outcome of patients due to the complications associated with fasciotomy. The purpose of the present study is to differentiate impending/incomplete compartment syndrome (ICS) from established acute compartment syndrome (ACS) in tibial fractures, and to assess the outcome of the fixation of the Ilizarov apparatus in patients with these fractures presenting with ICS, who were not submitted to fasciotomy. Methods After the establishment of the inclusion and exclusion criteria, 19 patients were included in the study from January 2007 to December 2017. All patients were male, with an average age of 42.3 ± 11.38 years. All of these patients were managed with Ilizarov ring fixation as per the medical and surgical protocol established in the present study. Results The average follow-up obtained for our 18 patients was of 47 ± 41.5 months, with one patient being lost to follow-up. The average time for ring application was of 3.7 ± 1.7 days. In total, 3 (16.7%) of these patients had nonunion. There were no soft-tissue or neurovascular complications in the immediate postoperative period. All of the patients eventually united and were independently mobile without any sequelae of compartment syndrome. Conclusion The Ilizarov ring external fixator can be used in the management of tibial fractures with ICS, avoiding fasciotomy with its various complications of infection and nonunion, resulting in fewer surgeries and faster rehabilitation. Surgeons should carefully differentiate ACS and ICS in these patients, as the clinical and functional results vary significantly. Unnecessary fasciotomies should be avoided.

10.
Cartilage ; 13(2_suppl): 808S-817S, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34528493

RESUMO

PURPOSE: Chondrocytes, isolated from articular cartilage, are routinely utilized in cell-based therapeutics for the treatment of cartilage pathologies. However, restoration of the biological tissue faces hindrance due to the formation of primarily fibrocartilaginous repair tissue. Chondroprogenitors have been reported to display superiority in terms of their chondrogenic potential and lesser proclivity for hypertrophy. In line with our recent results, comparing chondroprogenitors and chondrocytes, we undertook isolation of progenitors from the general pool of chondrocytes, based on surface marker expression, namely, CD166, CD34, and CD146, to eliminate off-target differentiation and generate cells of stronger chondrogenic potential. This study aimed to compare chondrocytes, chondroprogenitors, CD34-CD166+CD146+ sorted chondrocytes, and CD34-CD166+CD146- sorted chondrocytes. METHODS: Chondrocytes obtained from 3 human osteoarthritic knee joints were subjected to sorting, to isolate CD166+ and CD34- subsets, and then were further sorted to obtain CD146+ and CD146- cells. Chondrocytes and fibronectin adhesion-derived chondroprogenitors served as controls. Assessment parameters included reverse transcriptase polymerase chain reaction for markers of chondrogenesis and hypertrophy, trilineage differentiation, and total GAG/DNA content. RESULTS: Based on gene expression analysis, CD34-CD166+CD146+ sorted chondrocytes and chondroprogenitors displayed comparability and significantly higher chondrogenesis with a lower tendency for hypertrophy when compared to chondrocytes and CD34-CD166+CD146- sorted chondrocytes. The findings were also reiterated in multilineage potential differentiation with the 146+ subset and chondroprogenitors displaying lower calcification and chondroprogenitors displaying higher total GAG/DNA content compared to chondrocytes and 146- cells. CONCLUSION: This unique progenitor-like population based on CD34-CD166+CD146+ sorting from chondrocytes exhibits efficient potential for cartilage repair and merits further evaluation for its therapeutic application.


Assuntos
Antígenos CD34/imunologia , Antígenos CD/imunologia , Cartilagem Articular , Moléculas de Adesão Celular Neuronais/imunologia , Condrócitos , Proteínas Fetais/imunologia , Antígeno CD146/metabolismo , Diferenciação Celular , Condrócitos/metabolismo , Condrogênese/genética , Humanos
11.
J Bone Joint Surg Am ; 103(21): 2006-2013, 2021 11 03.
Artigo em Inglês | MEDLINE | ID: mdl-34138780

RESUMO

BACKGROUND: A subgroup of pertrochanteric fractures-namely, the AO/OTA 31A3 fracture-continues to be a difficult problem to treat, even with cephalomedullary nails. We present the results for 26 patients with a 31A3 fracture treated with the angled blade plate. METHODS: The records of 26 consecutive patients with a 31A3 fracture that was treated operatively with the angled blade plate device between 2007 and 2012 at our center were reviewed, and the patients were contacted for follow-up. The functional outcome (traumatic hip rating score) and radiographic outcome (the neck-shaft angle at the time of fixation and final follow-up) were obtained for 20 of the 26 patients at a minimum follow-up of 1 year. RESULTS: All 26 patients had primary surgery. At final review, 2 patients had died and 4 had been lost to follow-up. Of the 4 patients lost to follow-up, 2 had revision of the fixation with the angled blade plate. Of the 20 patients with follow-up, 1 had malreduction and implant failure but eventually had healing after revision of the fixation with the angled blade plate. The mean traumatic hip rating score at the time of follow-up was 50.0 with 4, 14, and 2 patients having excellent, good, and failed outcomes, respectively. The mean neck-shaft angle at the time of final union was 126.16°, which was an average of 4° less than that on the unaffected side. However, this did not correlate with functional outcome. There was no significant difference between the immediate postoperative and final neck-shaft angles. CONCLUSIONS: This study demonstrated that blade plate fixation for 31A3 fractures is associated with low rates of failure (15%), revision surgery (15%), and infection (15%), which are comparable with the results of nail fixation (range, 5% to 12%) and superior to those of sliding hip screw fixation. This large series demonstrates that the angled blade plate can be utilized for these complex fractures and should be part of the armamentarium for these injuries. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.


Assuntos
Placas Ósseas/efeitos adversos , Fixação Interna de Fraturas/efeitos adversos , Fraturas do Quadril/cirurgia , Infecção da Ferida Cirúrgica/epidemiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Pinos Ortopédicos/efeitos adversos , Pinos Ortopédicos/estatística & dados numéricos , Placas Ósseas/estatística & dados numéricos , Parafusos Ósseos/efeitos adversos , Parafusos Ósseos/estatística & dados numéricos , Estudos de Viabilidade , Feminino , Seguimentos , Fixação Interna de Fraturas/instrumentação , Fixação Interna de Fraturas/métodos , Consolidação da Fratura , Humanos , Masculino , Pessoa de Meia-Idade , Reoperação/estatística & dados numéricos , Estudos Retrospectivos , Infecção da Ferida Cirúrgica/etiologia , Resultado do Tratamento , Adulto Jovem
12.
Knee ; 30: 51-62, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33857741

RESUMO

BACKGROUND: Articular chondroprogenitors are a promising contender for cartilage repair due to their inherent nature which stands primed for chondrogenesis and minimal hypertrophic preponderance. Platelet rich plasma (PRP) has been extensively used for treating cartilage defects and osteoarthritis (OA), due to its chondro-inductive properties and abundant pool of growth factors. The aim of this study was to assess the efficacy of chondroprogenitors injected with PRP versus PRP alone in the healing of experimentally created early OA and osteochondral defects (OCD) in a rabbit model. METHODS: Adult New Zealand White male rabbits were used for cell and PRP isolation. Chondroprogenitors were isolated by fibronectin adhesion assay, labelled with iron oxide, characterized for surface markers, differential potential and expanded. PRP was isolated by double spin centrifugation using a TriCell kit. Study groups included (a) Monosodium iodoacetate induced early OA and (b) critical OCD. Following intervention (test arm: PRP+ chondroprogenitors and control arm: PRP), assessment was performed at 6- and 12-weeks which included histopathological examination and scoring (OARSI and Modified Wakitani score), immunohistochemistry analysis (Collagen type II and X) and synovial fluid S100A12 levels. RESULTS AND CONCLUSION: Comparable, evident healing was noticed in both test and control arms when the OA group samples were assessed at both time points. In the OCD group, PRP alone exhibited significantly better results than the test arm, although repair was notable in both interventions. Further evaluation of chondroprogenitors is required to assess their role as a standalone therapy and in combination with PRP to further cartilage regeneration.


Assuntos
Cartilagem Articular/fisiopatologia , Osteoartrite do Joelho/terapia , Plasma Rico em Plaquetas , Células-Tronco/citologia , Animais , Cartilagem Articular/citologia , Diferenciação Celular , Células Cultivadas , Condrogênese , Colágeno Tipo II/metabolismo , Modelos Animais de Doenças , Masculino , Osteoartrite do Joelho/induzido quimicamente , Coelhos , Proteína S100A12/metabolismo , Células-Tronco/fisiologia , Líquido Sinovial/metabolismo
13.
Nutrients ; 13(4)2021 Apr 13.
Artigo em Inglês | MEDLINE | ID: mdl-33924396

RESUMO

BACKGROUND: Few preclinical studies have shown that Knee osteoarthritis (KOA) is linked to gut microbiome dysbiosis and chronic inflammation. This pilot study was designed to look at the gut microbiome composition in KOA patients and normal individuals with or without vitamin D deficiency (VDD, serum vitamin D <30 ng/mL). METHODS: This pilot study was conducted prospectively in 24 participants. The faecal samples of all the participants were taken for DNA extraction. The V3-V4 region of 16s rRNA was amplified, and the library was prepared and sequenced on the Illumina Miseq platform. RESULTS: The mean (±SD) age was 45.5 (±10.2) years with no defined comorbidities. Of 447 total Operational Taxonomic Units (OTUs), a differential abundance of 16 nominally significant OTUs between the groups was observed. Linear discriminate analysis (LEfSe) revealed a significant difference in bacteria among the study groups. Pseudobutyrivibrio and Odoribacter were specific for VDD, while Parabacteroides, Butyricimonas and Gordonibacter were abundant in the KOA_VDD group, and Peptococcus, Intestimonas, Delftia and Oribacterium were abundant in the KOA group. About 80% of bacterial species were common among different groups and hence labelled as core bacterial species. However, the core microbiome of KOA and VDD groups were not seen in the KOA_VDD group, suggesting that these bacterial groups were affected by the interaction of the KOA and VDD factors. CONCLUSION: Parabacteroides, Butyricimonas, Pseudobutyrivibrio, Odoribacter and Gordonibacter are the predominant bacteria in vitamin D deficient patients with or without KOA. Together these results indicate an association between the gut microbiome, vitamin D and knee osteoarthritis.


Assuntos
Disbiose/complicações , Microbioma Gastrointestinal/imunologia , Osteoartrite do Joelho/imunologia , Deficiência de Vitamina D/imunologia , Adulto , DNA Bacteriano/isolamento & purificação , Disbiose/diagnóstico , Disbiose/imunologia , Disbiose/microbiologia , Fezes/microbiologia , Microbioma Gastrointestinal/genética , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Pessoa de Meia-Idade , Osteoartrite do Joelho/microbiologia , Filogenia , Projetos Piloto , RNA Ribossômico 16S/genética , Vitamina D/sangue , Deficiência de Vitamina D/sangue , Deficiência de Vitamina D/diagnóstico , Deficiência de Vitamina D/microbiologia
14.
J Clin Orthop Trauma ; 14: 22-28, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33717892

RESUMO

BACKGROUND: The ability to grow new cartilage remains the standard goal of any treatment strategy directed at cartilage repair. Chondroprogenitors have garnered interest due to their applicability in cell therapy. Pulsed electromagnetic field (PEMF) favors chondrogenesis by possible upregulation of genes belonging to TGFß superfamily. Since TGFß is implicated in chondrogenic signalling, the aim of the study was to evaluate the ability of PEMF to induce chondrogenesis via endogenous TGFß production in chondroprogenitors vs differentiation using chondrogenic medium inclusive of TGFß. METHODS: Chondroprogenitors were harvested from three non-diseased human knee joints via fibronectin assay. Passage 3 pellets were subjected to four different culture conditions: a) negative control contained chondrogenic medium without TGFß2, b) positive control contained medium with TGFß2, c) PEMF 1 contained medium of negative control plus single exposure to PEMF and d) PEMF 2 contained medium of negative control plus multiple exposures to PEMF. Following differentiation (day 21), pellets were assessed for gene expression of ACAN, SOX9, COL2A1, TGFß1, TGFß2, and TGFß3. Alcian blue staining to detect glycosaminoglycan deposition was also performed. Medium supernatant was used to detect endogenous latent TGF-ß1 levels using ELISA. RESULTS: All study arms exhibited comparable gene expression without any significant difference. Although positive control and PEMF study arms demonstrated notably better staining than negative control, the level of latent TGF-ß1 was seen to be significantly high in supernatant from positive control (P < 0.05) when compared to other groups. CONCLUSION: Our results indicate that PEMF induced chondrogenesis might involve other signalling molecules, which require further evaluation.

15.
Connect Tissue Res ; 62(4): 427-435, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-32406271

RESUMO

Purpose: Cartilage repair following trauma or degeneration is poor, making cell-based therapy an important avenue of treatment. Chondrocytes and mesenchymal stem cells have been extensively studied as potential candidates, although tendency toward hypertrophy and formation of mixed hyaline-fibrocartilage necessitates further optimization. Chondroprogenitors, isolated using fibronectin adhesion assay are reported to show reduced hypertrophy and enhanced chondrogenesis. Laminin, an essential component of extracellular matrix, has been shown to positively modulate chondrocyte proliferation, migration, and survival. The aim of our study was to evaluate the effect of laminin as a differential adhesion assay and obtain an enriched population of chondroprogenitors and assess its efficiency when compared to progenitors obtained via fibronectin.Materials and methods: Chondrocytes were isolated from three osteoarthritic knee joints and subjected to fibronectin and laminin adhesion to obtain chondroprogenitors. After expansion in culture, they were assessed for differences in their biological characteristics based on growth kinetics, surface marker expression, gene expression for assessing markers of chondrogenesis and hypertrophy, and potential for tri-lineage differentiation.Results: Our results showed that cells isolated by laminin and fibronectin both displayed comparable characteristics except in terms of proliferative potential (higher in laminin), gene expression of COL2A1 (lower in laminin) and trilineage potential where the laminin group showed higher osteogenic and adipogenic differentiation.Conclusion: This was the first attempt to successfully isolate human articular cartilage derived chondroprogenitor clones using laminin, which retained stem cell like characteristics. Further evaluation to optimize this method will help enhance chondroprogenitor characteristics, for use in cartilage repair.


Assuntos
Cartilagem Articular , Laminina , Condrogênese , Fibronectinas , Humanos , Hipertrofia
16.
Cartilage ; 13(2_suppl): 34S-52S, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-32840123

RESUMO

OBJECTIVE: Chondroprogenitors have recently gained prominence due to promising results seen in in vitro and animal studies as a potential contender in cell-based therapy for cartilage repair. Lack of consensus regarding nomenclature, isolation techniques, and expansion protocols create substantial limitations for translational research, especially given the absence of distinct markers of identification. The objective of this systematic review was to identify and collate information pertaining to hyaline cartilage-derived chondroprogenitors, with regard to their isolation, culture, and outcome measures. DESIGN: As per Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines, a web-based search of Scopus and PubMed databases was performed from January 2000 to May 2020, which yielded 509 studies. A total of 65 studies were identified that met the standardized inclusion criteria which comprised of, but was not limited to, progenitors derived from fibronectin adhesion, migrated subpopulation from explant cultures, and single-cell sorting. RESULT: Literature search revealed that progenitors demonstrated inherent chondrogenesis and minimal tendency for hypertrophy. Multiple sources also demonstrated significantly better outcomes that bone marrow-derived mesenchymal stem cells and comparable results to chondrocytes. With regard to progenitor subgroups, collated evidence points to better and consistent outcomes with the use of migratory progenitors when compared to fibronectin adhesion assay-derived progenitors, although a direct comparison between the two cell populations is warranted. CONCLUSION: Since chondroprogenitors exhibit favorable properties for cartilage repair, efficient characterization of progenitors is imperative, to complete their phenotypic profile, so as to optimize their use in translational research for neocartilage formation.


Assuntos
Cartilagem Articular , Células-Tronco Mesenquimais , Animais , Condrócitos , Condrogênese , Cartilagem Hialina
17.
J Clin Orthop Trauma ; 11(Suppl 4): S506-S511, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32774019

RESUMO

OBJECTIVES: Accurate diagnosis of osteoarthritis (OA) is the first important step in ensuring appropriate management of the disease. A multitude of tests involving assessment of biomarkers help in assessment of severity and grading of osteoarthritic damage. However, most tests are time consuming and are limited by the paucity in synovial fluid volume. In majority of OA effusions, calcium containing crystals are found. The aim of our study was to evaluate whether a correlation existed between the amount of calcium containing crystals present in synovial fluid and severity scoring of OA to propose a quick and inexpensive technique for disease assessment. MATERIALS AND METHODS: Monosodium-iodoacetate was used to induce high- and low-grade knee OA in adult New Zealand white rabbits (n = 6 joint each group). At 16 weeks, synovial fluid and joints were harvested for histopathological analysis. OA grading was established based on OARSI scoring. Synovial fluid calcium crystal count was assessed by light microscopy (Alizarin red) and confirmed by Fluo-4, AM imaging and polarized microscopy. Statistical analysis was performed using unpaired Student t-test and Pearson correlation. RESULTS AND CONCLUSION: The clumps counted in low-grade OA were significantly lower than high-grade OA, in addition to showing a positive correlation (coefficient: 0.65; P=0.021) between calcium crystal count and the grade of OA created. Fluo-4, AM staining, and polarized microscopy were indicative of calcium pyrophosphate dihydrate crystals. This is the first study to suggest that Alizarin red could serve as an effective and rapid, bed-side method for screening and assessing disease progression.

18.
Acta Histochem ; 122(6): 151588, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32778244

RESUMO

PURPOSE: Cell based therapy is constantly underway since regeneration of genuine hyaline cartilage is under par. Much attention has been afforded to chondroprogenitors recently, as an alternative cell substitute for cartilage repair. Although single source derivation of chondrocytes and chondroprogenitors is advantageous, lack of a characteristic differentiating marker obscures clear identification, which is essential to create a biological profile and is also required to assess cell type superiority for cartilage repair. METHODS: Cells obtained from three non-diseased/osteoarthritic human knee joints each, were expanded in culture up to passage 10. Characterization studies were performed using flow cytometry; gene expression was studied using RT-PCR; growth kinetics and tri-lineage differentiation was also studied to construct a better profile of chondroprogenitors as well as chondrocytes. RESULTS AND CONCLUSION: Our results showed that both cell populations exhibited similar cell surface characteristics except for non-diseased chondroprogenitors, which showed markedly low expression of CD34 and high expression of CD166. Trilineage data was suggestive of multilineage potential for both cell types with chondroprogenitors showing notably higher glycosaminoglycan and lower calcified matrix deposition. Data acquired from this study aided in describing cellular behavior of human articular cartilage derived chondroprogenitors in conditions not reported earlier. Our comparative analysis suggests that sorting based on a combination of markers (CD34- and CD166+) would yield a population of cells with minimal contamination by chondrocytes, which may provide translatable results in terms of enhanced chondrogenesis and reduced hypertrophy; both indispensable for the field of cartilage regeneration.


Assuntos
Terapia Baseada em Transplante de Células e Tecidos/métodos , Condrócitos/metabolismo , Cartilagem Articular/metabolismo , Condrogênese/fisiologia , Fibronectinas/metabolismo , Humanos , Articulação do Joelho/citologia , Osteoartrite/metabolismo
19.
Nanomedicine ; 29: 102232, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32562860

RESUMO

Healing or reconstruction of critical-sized bone defects is still challenging in orthopaedic practice. In this study, we developed a new approach to control the degradation and improve the bone regeneration of the AZ31 magnesium substrate, fabricated as mesh cage implants. Subsequently, bilayer nanocomposite coating was carried out using polycaprolactone (PCL) and nano-hydroxyapatite (nHA) by dip-coating and electrospinning. Lastly, the healing capacity of the implants was studied in New Zealand White (NZW) rabbit critical-sized femur bone defects. X-ray analysis showed the coated implant group bridged and healed the critical defects 100% during four weeks of post-implantation. Micro-computed tomography (Micro-CT) study showed higher total bone volume (21.10%), trabecular thickness (0.73), and total porosity (85.71%) with bilayer coated implants than uncoated. Our results showed that nanocomposite coated implants controlled the in vivo degradation and improved bioactivity. Hence, the coated implants can be used as a promising bioresorbable implant for critical segmental bone defect repair applications.


Assuntos
Regeneração Óssea/efeitos dos fármacos , Materiais Revestidos Biocompatíveis/farmacologia , Nanoestruturas/química , Próteses e Implantes , Ligas/química , Ligas/farmacologia , Animais , Durapatita/farmacologia , Fêmur/efeitos dos fármacos , Fêmur/crescimento & desenvolvimento , Humanos , Magnésio/química , Magnésio/farmacologia , Nanocompostos/química , Nanocompostos/uso terapêutico , Poliésteres/química , Poliésteres/farmacologia , Coelhos , Microtomografia por Raio-X
20.
Knee ; 27(3): 833-837, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32317141

RESUMO

BACKGROUND: Cell-based therapy in cartilage repair can benefit from the use of chondroprogenitors; a cell type classified as mesenchymal stem cells, demonstrating reduced hypertrophy. Fibronectin, routinely used to isolate chondroprogenitors, classically binds to α5ß1 integrins (CD49e + CD29), of which CD49e is said to be highly expressed in progenitors. The aim of our study was to assess the specificity of CD49e as a distinguishing marker for chondroprogenitors; because studies report low expression in fresh chondrocytes (FCs), but recent conflicting data has exhibited incremental expression of CD49e in cultured chondrocytes. METHODS: FCs were isolated from three human osteoarthritic knee joints and CD49e- cells (sorted by flow cytometry) were cultured in adherent and non-adherent conditions and reassessed for CD49e and CD29 at multiple time points. Colony-forming efficiency (CFE) following fibronectin adhesion assay was calculated for FC, CD49e+ and CD49e- cells. RESULTS: A statistically significant increase in CD49e and CD29 expression was seen in both adherent and non-adherent cultures of CD49e- cells (P < 0.01), as early as 24 h. All groups grew clonally and CFE was similar without any significant difference. CD49e- chondrocytes turned positive when cultured, possibly due to an inherent phenotypic drift, seen after release from cartilage and not because of plastic adherence or chondroprogenitor overgrowth, as non-adherent cultures also showed high expression. CONCLUSIONS: As the specificity of CD49e is questionable, there is a pressing need for a specific differentiating marker, to isolate a pure population of chondroprogenitors, as this cell type shows inherent chondrogenesis and reduced hypertrophy, both requisites for cartilage repair.


Assuntos
Cartilagem Articular/patologia , Condrócitos/citologia , Integrina alfa1/metabolismo , Osteoartrite do Joelho/metabolismo , Adulto , Biomarcadores/metabolismo , Cartilagem Articular/metabolismo , Diferenciação Celular , Células Cultivadas , Condrogênese , Humanos , Articulação do Joelho , Masculino , Células-Tronco Mesenquimais , Pessoa de Meia-Idade , Osteoartrite do Joelho/patologia
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