Spinopelvic interactions in total hip arthroplasty: 295 patients followed for a minimum follow-up of 10 years.

BACKGROUND: Concurrent spinal pathology is frequent in patients undergoing total hip arthroplasty (THA). In this study we examined whether spinopelvic interactions affect THA outcomes at a minimum follow-up of 10 years. PATIENTS AND METHODS: 295 patients with a mean age of 63.3 (range 56‒80) years receiving a THA between 2006 and 2009 were assessed. Of these, 195 had mild lumbar disc degeneration and 100 had advanced lumbar spondylosis. We analysed the changes in the Harris Hip Score (HHS) and the survival rate for postoperative low back pain (LBP) and dislocation. Changes in acetabular component position, sacro-femoral-pubic (SFP) and pelvic obliquity (PO) angles were assessed with radiological images. RESULTS: The mean HHS was lower in female patients (p = 0.009), patients >65 years of age (p < 0.001) and those with advanced lumbar spondylosis (p = 0.002). 52 (71.2%) of the patients reporting preoperative LBP experienced improvement after THA while 47 (21.1%) of those without preoperative LBP postoperatively reported new onset LBP. Female patients (p = 0.025; hazard ratio [HR]: 1.831; 95% CI, 1.081-3.101) and those with preoperative LBP (p = 0.007; HR 2.068; 95% CI, 1.221-3.504) were at a higher risk of developing postoperative LBP at 10 years. 4 out of 9 THA dislocations were late and had shown decreasing SFP angle values over time. Acetabular component inclination and anteversion angles increased over time, whereas the SFP angle was associated with sex and age and the PO angle with age and the severity of any preoperative lumbar degeneration. CONCLUSIONS: Concurrent spinal pathology influences THA outcomes at a minimum follow-up of 10 years. Sex, age, and associated lumbar degeneration can affect clinical and radiological changes over time. A decrease in SFP angle values over time was found in patients sustaining late dislocation.

Long-term results of total hip arthroplasty in severe congenital hip disease: A ten- to 20-year follow-up study.

INTRODUCTION: The impact of bone deformities, previous surgeries, and the surgical technique in total hip arthroplasty (THA) for congenital dislocation of the hip (CDH) at a long-term has not been clearly defined yet. In this single-centre observational study we sought to assess patients undergoing THA due to osteoarthritis secondary to severe CDH with low or high dislocation ten- to 20-years after surgery. To determine this purpose, we assessed: (1) THA-related complications and reoperations; (2) the clinical outcome, patients' satisfaction and radiological results; and (3) the possible risk factors for reoperation with particular attention to the surgical technique and the influence of prior surgeries. HYPOTHESIS: We hypothesized that an anatomical reconstruction of the hip would decrease the reoperations rates in patients undergoing THA with severe CDH. METHODS: Seventy-five patients (85 hips) operated between 1999 and 2012 at our large tertiary hospital were analyzed. Fifty-six hips were diagnosed as low dislocation (group 1) and 29 hips as high dislocation (group 2). The existence of prior surgeries was frequent: group 1, pelvic osteotomies 6 hips, femoral osteotomies 7, tectoplasty (shelf) 6, resection arthroplasty 1 and lowering of the greater trochanter 1; group 2 included pelvic osteotomies 10 hips, femoral osteotomies 10, and a femoral lengthening 2. The number of additional procedures during THA was: group 1 (19/56 [34%]), acetabular roof bone autograft 8 hips, acetabular medial wall autograft 2, hardware removal 6, extended femoral osteotomy 2, and a femoral shortening 1; group 2 (20/29 [69%]), acetabular bone autograft 12 hips, medial autograft 1, hardware removal 1, extended femoral osteotomy 2, and a femoral shortening 4 hips. The clinical and the radiological analysis were compared in both groups for a minimum follow-up of ten years. Cox regression models were used to detect risk factors for reoperation. RESULTS: Nine patients (13.8%) required reoperation for the following reasons: cup loosening (5 hips), periprosthetic femoral fracture (3) and stem loosening (1); seven had prior surgeries. The 12-year survival rate for reoperation for any reason was 96.3% (95% confidence interval [CI] 91.2-100) in group 1 and 75.7% (95% CI, 65.8-90.8) in group 2 (p=0.003). Patients with high dislocation (p=0.02, hazard ratio [HR]: 6.25, 95% CI, 1.26-30.9) and those with an acetabular component inclination placed out of the target zone between 35° and 50° (p=0.03, HR: 4.27, 95% CI, 1.13-16.1) had a higher risk of reoperation. DISCUSSION: An optimal placement of the acetabular component decreased the reoperation rates in patients undergoing THA for severe CDH. Hips with high dislocation and the existence of prior surgery can affect THA implantation. LEVEL OF EVIDENCE: III; retrospective; comparative.

The Epidemic of Internalizing Problems Among Latinx Adolescents Before and During the Coronavirus 2019 Pandemic.

OBJECTIVE: Latinx youth exhibit disproportionately higher internalizing symptoms than their peers from other racial/ethnic groups. This study compares depression and anxiety symptoms between referred students of Latinx and non-Latinx backgrounds before and during the COVID-19 pandemic and examines key determinants within the Latinx sample. METHOD: Data are analyzed from four academic years - two before and two during the pandemic - from 1220 5th through 8th grade students (Mage = 12.1; 59.6% female; 59.9% Latinx or mixed-Latinx) referred for services across 59 Chicago Public School District (CPS) elementary schools. Using the Children's Depression Inventory (CDI) and the Revised Child Anxiety and Depression Scale (RCADS), mean scores and risk levels for depression, social anxiety, and generalized anxiety are examined. RESULTS: Higher internalizing risk and comorbidity rates were found in the second year of the pandemic, compared to pre-pandemic levels. Latinx students reported higher depression, social anxiety, and generalized anxiety symptoms than non-Latinx students. During the pandemic, more Latinx students were classified as having comorbid depression and anxiety, and scored in the clinical range for depression, generalized anxiety, and social anxiety than non-Latinx students. Within the Latinx sample, girls and gender non-conforming students reported the highest maladjustment. CONCLUSIONS: Results highlight the pressing need to examine the long-term impact of COVID-19 on the mental health of Latinx children and adolescents, and to address their internalizing problems.

Sports Participation, Frequency, and Competence Differentially Impact Youth Depressive, Anxious, and Somatic Symptoms: Gender, Neighborhood, and Sports Type Effects.

Mental health problems are prevalent in adolescence, but sports participation may offer mental health benefits through this developmental period and beyond. Characteristics of sports participation including perceived frequency and competence may differentially predict adolescent depressive, anxious, and somatic symptoms over time and results may further vary according to gender, neighborhood context, and type of sport engagement. Data were collected at two time-points six months apart from an ethnically diverse sample of adolescents (N = 183, female = 51%). Youth sports participation and symptoms were measured using the Youth Self-Report (YSR; Achenbach & Rescorla, 2001). Path analyses were used to test for main and moderating effects of sports on symptoms. Results showed that categorical sports participation did not prospectively predict any type of internalizing symptoms, but perceived frequency and competence did. Competence predicted lower levels of symptoms while frequency predicted higher levels of symptoms. These results were further moderated by gender, neighborhood, and sport type such that frequency and competence predicted symptoms for girls and for youth in more resourced neighborhoods and who participated in team sports. These findings highlight the impact that sports participation can have on adolescent mental health in an ethnically diverse sample of urban youth.

Selection and Optimization of a Bioink Based on PANC-1- Plasma/Alginate/Methylcellulose for Pancreatic Tumour Modelling.

3D bioprinting involves using bioinks that combine biological and synthetic materials. The selection of the most appropriate cell-material combination for a specific application is complex, and there is a lack of consensus on the optimal conditions required. Plasma-loaded alginate and alginate/methylcellulose (Alg/MC) inks were chosen to study their viscoelastic behaviour, degree of recovery, gelation kinetics, and cell survival after printing. Selected inks showed a shear thinning behavior from shear rates as low as 0.2 s-1, and the ink composed of 3% w/v SA and 9% w/v MC was the only one showing a successful stacking and 96% recovery capacity. A 0.5 × 106 PANC-1 cell-laden bioink was extruded with an Inkredible 3D printer (Cellink) through a D = 410 µm tip conical nozzle into 6-well culture plates. Cylindrical constructs were printed and crosslinked with CaCl2. Bioinks suffered a 1.845 Pa maximum pressure at the tip that was not deleterious for cellular viability. Cell aggregates can be appreciated for the cut total length observed in confocal microscopy, indicating a good proliferation rate at different heights of the construct, and suggesting the viability of the selected bioink PANC-1/P-Alg3/MC9 for building up three-dimensional bioprinted pancreatic tumor constructs.

New polyethylenes in total hip arthroplasty : a 20- to 22-year follow-up study.

AIMS: A significant reduction in wear at five and ten years was previously reported when comparing Durasul highly cross-linked polyethylene with nitrogen-sterilized Sulene polyethylene in total hip arthroplasty (THA). We investigated whether the improvement observed at the earlier follow-up continued, resulting in decreased osteolysis and revision surgery rates over the second decade. METHODS: Between January 1999 and December 2001, 90 patients underwent surgery using the same acetabular and femoral components with a 28 mm metallic femoral head and either a Durasul or Sulene liner. A total of 66 hips of this prospective randomized study were available for a minimum follow-up of 20 years. The linear femoral head penetration rate was measured at six weeks, one year, and annually thereafter, using the Dorr method on digitized radiographs with a software package. RESULTS: In the Durasul group, no patients underwent revision due to loosening or showed radiological evidence of osteolysis. In the Sulene group, four patients (four hips) were revised due to femoral component loosening. The 20-year cumulative failure incidence in the presence of the competing event of death for revision surgery was 4.5% (95% confidence interval (CI) 0.8 to 13.6) in the Durasul group, and 8.9% (95% CI 2.8 to 19.5) in the Sulene group. The mean wear one year after surgery was 0.09 mm (SD 0.007) in the Durasul group and 0.24 (SD 0.015) in the Sulene group (p < 0.001). From one to 20 years after surgery, the mean total penetration was 0.32 mm (SD 0.045) in the Durasul group and 1.07 mm (SD 0.13) in the Sulene group (p < 0.001). Mean femoral head penetration at 20 years was approximately 70.0% less in the Durasul group than the Sulene group. CONCLUSION: The significant reduction in femoral head penetration obtained with the Durasul compared with Sulene in uncemented THA resulted in lower osteolysis and revision rates after 20 years.Cite this article: Bone Joint J 2022;104-B(9):1032-1038.

Polylactide, Processed by a Foaming Method Using Compressed Freon R134a, for Tissue Engineering.

Fabricating polymeric scaffolds using cost-effective manufacturing processes is still challenging. Gas foaming techniques using supercritical carbon dioxide (scCO2) have attracted attention for producing synthetic polymer matrices; however, the high-pressure requirements are often a technological barrier for its widespread use. Compressed 1,1,1,2-tetrafluoroethane, known as Freon R134a, offers advantages over CO2 in manufacturing processes in terms of lower pressure and temperature conditions and the use of low-cost equipment. Here, we report for the first time the use of Freon R134a for generating porous polymer matrices, specifically polylactide (PLA). PLA scaffolds processed with Freon R134a exhibited larger pore sizes, and total porosity, and appropriate mechanical properties compared with those achieved by scCO2 processing. PLGA scaffolds processed with Freon R134a were highly porous and showed a relatively fragile structure. Human mesenchymal stem cells (MSCs) attached to PLA scaffolds processed with Freon R134a, and their metabolic activity increased during culturing. In addition, MSCs displayed spread morphology on the PLA scaffolds processed with Freon R134a, with a well-organized actin cytoskeleton and a dense matrix of fibronectin fibrils. Functionalization of Freon R134a-processed PLA scaffolds with protein nanoparticles, used as bioactive factors, enhanced the scaffolds' cytocompatibility. These findings indicate that gas foaming using compressed Freon R134a could represent a cost-effective and environmentally friendly fabrication technology to produce polymeric scaffolds for tissue engineering approaches.

Osteoblast function in patients with idiopathic osteonecrosis of the femoral head : implications for a possible novel therapy.

AIMS: To investigate whether idiopathic osteonecrosis of the femoral head (ONFH) is related to impaired osteoblast activities. METHODS: We cultured osteoblasts isolated from trabecular bone explants taken from the femoral head and the intertrochanteric region of patients with idiopathic ONFH, or from the intertrochanteric region of patients with osteoarthritis (OA), and compared their viability, mineralization capacity, and secretion of paracrine factors. RESULTS: Osteoblasts from the intertrochanteric region of patients with ONFH showed lower alkaline phosphatase (ALP) activity and mineralization capacity than osteoblasts from the same skeletal site in age-matched patients with OA, as well as lower messenger RNA (mRNA) levels of genes encoding osteocalcin and bone sialoprotein and higher osteopontin expression. In addition, osteoblasts from patients with ONFH secreted lower osteoprotegerin (OPG) levels than those from patients with OA, resulting in a higher receptor activator of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) ligand (RANKL)-to-OPG ratio. In patients with ONFH, osteoblasts from the femoral head showed reduced viability and mineralized nodule formation compared with osteoblasts from the intertrochanteric region. Notably, the secretion of the pro-resorptive factors interleukin-6 and prostaglandin E2 as well as the RANKL-to-OPG ratio were markedly higher in osteoblast cultures from the femoral head than in those from the intertrochanteric region. CONCLUSION: Idiopathic ONFH is associated with a reduced mineralization capacity of osteoblasts and increased secretion of pro-resorptive factors. Cite this article: Bone Joint Res 2021;10(9):619-628.

Impaction bone grafting in hip re-revision surgery.

AIMS: Bone stock restoration of acetabular bone defects using impaction bone grafting (IBG) in total hip arthroplasty may facilitate future re-revision in the event of failure of the reconstruction. We hypothesized that the acetabular bone defect during re-revision surgery after IBG was smaller than during the previous revision surgery. The clinical and radiological results of re-revisions with repeated use of IBG were also analyzed. METHODS: In a series of 382 acetabular revisions using IBG and a cemented component, 45 hips (45 patients) that had failed due to aseptic loosening were re-revised between 1992 and 2016. Acetabular bone defects graded according to Paprosky during the first and the re-revision surgery were compared. Clinical and radiological findings were analyzed over time. Survival analysis was performed using a competing risk analysis. RESULTS: Intraoperative bone defect during the initial revision included 19 Paprosky type IIIA and 29 Paprosky type IIIB hips; at re-revision, seven hips were Paprosky type II, 27 type IIIA and 11 were type IIIB (p = 0.020). The mean preoperative Harris Hip Score was 45.4 (SD 6.4), becoming 80.7 (SD 12.7) at the final follow-up. In all, 12 hips showed radiological migration of the acetabular component, and three required further revision surgery. The nine-year cumulative failure incidence (nine patients at risk) of the acetabular component for further revision surgery was 9.6% (95% confidence interval (CI) 2.9 to 21.0) for any cause, and 7.5% (95% CI 1.9 to 18.5) for aseptic loosening. Hips with a greater hip height had a higher risk for radiological migration (odds ratio 1.09, 95% CI 1.02 to 1.17; p = 0.008). CONCLUSION: Bone stock restoration can be obtained using IBG in revision hip surgery. This technique is also useful in re-revision surgery; however, a better surgical technique including a closer distance to hip rotation centre could decrease the risk of radiological migration of the acetabular component. A longer follow-up is required to assess potential fixation deterioration. Cite this article: Bone Joint J 2021;103-B(3):492-499.

Vitamin B9 derivatives as carriers of bioactive cations for musculoskeletal regeneration applications: Synthesis, characterization and biological evaluation.

The development of new drugs for musculoskeletal regeneration purposes has attracted much attention in the last decades. In this work, we present three novel vitamin B9 (folic acid)-derivatives bearing divalent cations (ZnFO, MgFO and MnFO), providing their synthesis mechanism and physicochemical characterization. In addition, a strong emphasis has been placed on evaluating their biological properties (along with our previously reported SrFO) using human mesenchymal stem cells (hMSC). In all the cases, pure folate derivatives (MFOs) with a bidentate coordination mode between the metal and the folate anion, and a 1:1 stoichiometry, were obtained in high yields. A non-cytotoxic dose of all the MFOs (50 µg/mL) was demonstrated to modulate by their own the mRNA profiles towards osteogenic-like or fibrocartilaginous-like phenotypes in basal conditions. Moreover, ZnFO increased the alkaline phosphatase activity in basal conditions, while both ZnFO and MnFO increased the matrix mineralization degree in osteoinductive conditions. Thus, we have demonstrated the bioactivity of these novel compounds and the suitability to further studied them in vivo for musculoskeletal regeneration applications.

Wear of hip prostheses increases serum IGFBP-1 levels in patients with aseptic loosening.

The biological mechanisms involved in aseptic loosening include inflammation-associated and bone resorption-associated processes. Coordinated cellular actions result in biochemical imbalances with devastating consequences for the joint. Given that this condition is not known for showing systemic signs, we investigated whether circulating levels of inflammation-related proteins are altered in patients with aseptic loosening. Our study included 37 patients who underwent revision surgery due to hip osteolysis and aseptic loosening and 31 patients who underwent primary total hip arthroplasty. Using antibody arrays, we evaluated the serum levels of 320 proteins in four patients from each group. The results showed differences in insulin-like growth factor-binding protein 1 (IGFBP-1) concentrations, which we then quantified using enzyme-linked immunosorbent assay tests in all study patients. The results confirmed that serum IGFBP-1 concentrations were higher in the revision surgery patients than in the hip arthroplasty patients. In vitro studies showed that exposure of human osteoblasts to titanium particles induced an IGFBP-1 release that further increased when exposure to particles was performed in media conditioned by human M1 macrophages. These findings suggest that elevated serum IGFBP-1 levels in patients with aseptic loosening can arise from increased local IGFBP-1 production in the inflammatory environment of the periprosthetic bed.

Parents report fewer executive functioning problems and repetitive behaviors in young dual-language speakers with autism.

More dual language learners (DLLs) are being identified early with autism spectrum disorder (ASD). However, many families are still being advised against dual language exposure, despite a lack of evidence of negative impacts on language development in ASD. Research in typically developing children has noted advantages for bilinguals in domains such as executive functioning and social skills, but less is known about the effects in ASD. The present study evaluated differences in executive functioning and social communication in young children (n = 55) with ASD. Dual-language learners with ASD had significantly fewer parent reported executive functioning problems and repetitive behaviors; parent-reported social communication skills were generally comparable across groups. Our findings indicate that the bilingual advantage in executive functioning may extend to children with neurodevelopmental conditions.

Influence of inflammatory conditions provided by macrophages on osteogenic ability of mesenchymal stem cells.

BACKGROUND: The mechanisms by which macrophage phenotype contributes to mesenchymal stem cells (MSC)-mediated bone repair remain unclear. In this work, we investigated the influence of factors released by human macrophages polarized to a pro-inflammatory or an anti-inflammatory phenotype on the ability of human MSC to attach, migrate, and differentiate toward the osteoblastic lineage. We focused on the role of TNF-α and IL-10, key pro-inflammatory and anti-inflammatory cytokines, respectively, in regulating MSC functions. METHODS: MSC were treated with media conditioned by pro-inflammatory or anti-inflammatory macrophages to study their influence in cell attachment, migration, and osteogenic differentiation. The involvement of TNF-α and IL-10 in the regulation of MSC functions was investigated using neutralizing antibodies and recombinant cytokines. RESULTS: Treatment of MSC with media conditioned by pro-inflammatory or anti-inflammatory macrophages promoted cell elongation and enhanced MSC ability to attach and migrate. These effects were more noticeable when MSC were treated with media from pro-inflammatory macrophages. Interestingly, MSC osteogenic activity was enhanced by factors released by anti-inflammatory macrophages, but not by pro-inflammatory macrophages. Significant IL-10 levels originated from anti-inflammatory macrophages enhanced MSC osteogenesis by increasing ALP activity and mineralization in MSC layers cultured under osteogenic conditions. Moreover, macrophage-derived IL-10 regulated the expression of the osteogenic markers RUNX2, COL1A1, and ALPL. Notably, low TNF-α levels secreted by anti-inflammatory macrophages increased ALP activity in differentiating MSC whereas high TNF-α levels produced by pro-inflammatory macrophages had no effects on osteogenesis. Experiments in which MSC were treated with cytokines revealed that IL-10 was more effective in promoting matrix maturation and mineralization than TNF-α. CONCLUSIONS: Factors secreted by pro-inflammatory macrophages substantially increased MSC attachment and migration whereas those released by anti-inflammatory macrophages enhanced MSC osteogenic activity as well as cell migration. IL-10 was identified as an important cytokine secreted by anti-inflammatory macrophages that potentiates MSC osteogenesis. Our findings provide novel insights into how environments provided by macrophages regulate MSC osteogenesis, which may be helpful to develop strategies to enhance bone regeneration.

Glycerylphytate compounds with tunable ion affinity and osteogenic properties.

Phytic acid (PA) is a natural-occurring antioxidant, which plays an important role in many biological processes. PA is recognized as a potent inhibitor of lipid peroxidation because of its high affinity to multivalent cations, and it can play a role in osteogenic processes. However, its powerful chelating capacity is controversial because it can lead to a severe reduction of mineral availability in the organism. For this reason, compounds with beneficial biological properties of PA, but a modular ion binding capacity, are of high interest. In this work, we report the synthesis and physicochemical characterization of two hydroxylic derivatives of PA, named glycerylphytates (GPhy), through a condensation reaction of PA with glycerol (G). Both derivatives present antioxidant properties, measured by ferrozine/FeCl2 method and chelating activity with calcium ions depending on the content of glyceryl groups incorporated. Besides, the hydroxylic modification not only modulates the ion binding affinity of derivatives but also improves their cytocompatibility in human bone marrow mesenchymal cells (MSCs). Furthermore, GPhy derivatives display osteogenic properties, confirmed by COL1A and ALPL expression depending on composition. These positive features convert GPhy compounds into potent alternatives for those skeletal diseases treatments where PA is tentatively applied.

Immunoregulatory potential of mesenchymal stem cells following activation by macrophage-derived soluble factors.

BACKGROUND: Immunoregulatory capacity of mesenchymal stem cells (MSC) is triggered by the inflammatory environment, which changes during tissue repair. Macrophages are essential in mediating the inflammatory response after injury and can adopt a range of functional phenotypes, exhibiting pro-inflammatory and anti-inflammatory activities. An accurate characterization of MSC activation by the inflammatory milieu is needed for improving the efficacy of regenerative therapies. In this work, we investigated the immunomodulatory functions of MSC primed with factors secreted from macrophages polarized toward a pro-inflammatory or an anti-inflammatory phenotype. We focused on the role of TNF-α and IL-10, prototypic pro-inflammatory and anti-inflammatory cytokines, respectively, as priming factors for MSC. METHODS: Secretion of immunoregulatory mediators from human MSC primed with media conditioned by human macrophages polarized toward a pro-inflammatory or an anti-inflammatory phenotype was determined. Immunomodulatory potential of primed MSC on polarized macrophages was studied using indirect co-cultures. Involvement of TNF-α and IL-10 in priming MSC and of PGE2 in MSC-mediated immunomodulation was investigated employing neutralizing antibodies. Collagen hydrogels were used to study MSC and macrophages interactions in a more physiological environment. RESULTS: Priming MSC with media conditioned by pro-inflammatory or anti-inflammatory macrophages enhanced their immunomodulatory potential through increased PGE2 secretion. We identified the pro-inflammatory cytokine TNF-α as a priming factor for MSC. Notably, the anti-inflammatory IL-10, mainly produced by pro-resolving macrophages, potentiated the priming effect of TNF-α. Collagen hydrogels acted as instructive microenvironments for MSC and macrophages functions and their crosstalk. Culturing macrophages on hydrogels stimulated anti-inflammatory versus pro-inflammatory cytokine secretion. Encapsulation of MSC within hydrogels increased PGE2 secretion and potentiated immunomodulation on macrophages, attenuating macrophage pro-inflammatory state and sustaining anti-inflammatory activation. Priming with inflammatory factors conferred to MSC loaded in hydrogels greater immunomodulatory potential, promoting anti-inflammatory activity of macrophages. CONCLUSIONS: Factors secreted by pro-inflammatory and anti-inflammatory macrophages activated the immunomodulatory potential of MSC. This was partially attributed to the priming effect of TNF-α and IL-10. Immunoregulatory functions of primed MSC were enhanced after encapsulation in hydrogels. These findings may provide insight into novel strategies to enhance MSC immunoregulatory potency.

Publisher Correction: Paracrine interactions between mesenchymal stem cells and macrophages are regulated by 1,25-dihydroxyvitamin D3.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Publisher Correction: Paracrine interactions between mesenchymal stem cells and macrophages are regulated by 1,25-dihydroxyvitamin D3.

A correction to this article has been published and is linked from the HTML version of this paper. The error has been fixed in the paper.

Paracrine interactions between mesenchymal stem cells and macrophages are regulated by 1,25-dihydroxyvitamin D3.

Mesenchymal stem cells (MSC) modulate the macrophage-mediated inflammatory response through the secretion of soluble factors. In addition to its classical effects on calcium homeostasis, 1,25-dihydroxyvitamin D3 (1,25D3) has emerged as an important regulator of the immune system. The present study investigates whether 1,25D3 modulates the paracrine interactions between MSC and macrophages. 1,25D3 stimulated MSC to produce PGE2 and VEGF and regulated the interplay between macrophages and MSC toward reduced pro-inflammatory cytokine production. Conditioned media (CM) from co-cultures of macrophages and MSC impaired MSC osteogenesis. However, MSC cultured in CM from 1,25D3-treated co-cultures showed increased matrix maturation and mineralization. Co-culturing MSC with macrophages prevented the 1,25D3-induced increase in RANKL levels, which correlated with up-regulation of OPG secretion. MSC seeding in three-dimensional (3D) substrates potentiated their immunomodulatory effects on macrophages. Exposure of 3D co-cultures to 1,25D3 further reduced the levels of soluble factors related to inflammation and chemotaxis. As a consequence of 1,25D3 treatment, the recruitment of monocytes toward CM of 3D co-cultures decreased, while the osteogenic maturation of MSC increased. These data add new insights into the pleiotropic effects of 1,25D3 on the crosstalk between MSC and macrophages and highlight the role of the hormone in bone regeneration.

Substrate Microarchitecture Shapes the Paracrine Crosstalk of Stem Cells with Endothelial Cells and Osteoblasts.

We examined the hypothesis that substrate microarchitecture regulates the crosstalk between human mesenchymal stem cells (hMSC) and cell types involved in bone regeneration. Compared with polyester flat substrates having uniformly distributed homogenous pores (2D), three-dimensional polystyrene substrates with randomly oriented and interconnected pores of heterogeneous size (3D) stimulated the stromal secretion of IGF-1 while lessened the production of VEGFR-1, MCP-1 and IL-6. The medium conditioned by hMSC cultured in 3D substrates stimulated tube formation by human endothelial cells (hEC) to a higher extent than medium from 2D cultures. 3D co-cultures of hMSC and hEC contained higher secreted levels of IGF-1, EGF and FGF-2 than 2D co-cultures, resulting in increased hEC proliferation and migration. Substrate microarchitecture influenced the secretion of factors related to bone remodeling as the ratio RANKL to OPG, and the levels of M-CSF and IL-6 were higher in 3D co-cultures of hMSC and human osteoblasts (hOB) than in 2D co-cultures. Cytokine microenvironment in 3D co-cultures stimulated osteoblast matrix reorganization while demoted the late steps of osteoblastic maturation. Altogether, data in this study may unveil a new role of scaffold microarchitecture during bone regeneration, as modulator of the paracrine relationships that hMSC establish with hEC and hOB.

Bioactivity of dexamethasone-releasing coatings on polymer/magnesium composites.

We developed biodegradable polymeric coatings loaded with increasing amounts of dexamethasone on composites based on polylactic acid and Mg particles for bone repair. Incorporation of Mg particles into the polymeric matrix improves the compressive behaviour of the polymer. Mg-containing composites release Mg2+ ions into the culture medium and improve mesenchymal stem cell (MSC) viability, enhance their osteogenic potential and promote the release of angiogenic factors. Dexamethasone-loaded coatings deposited on composites delay Mg2+ ion dissolution while releasing controlled amounts of the drug, which are highly dependent on initial payload. Release kinetic of dexamethasone from the coatings exhibits a fast initial release of the drug followed by a slower secondary release. Bioactivity of the released dexamethasone was explored by monitoring dose-dependent responses of MSCs and macrophages. Biological effects exerted by the released drug are similar to those observed in cells treated with solutions of the glucocorticoid, indicating that the method employed for inclusion of dexamethasone into the coatings does not impair its bioactive behaviour. Culturing MSCs on dexamethasone-releasing coatings enhances extracellular matrix production and initial induction to osteogenic commitment as a function of drug payload. Dexamethasone incorporated into the coatings presents anti-inflammatory activity, as shown by the decrease in the production of cytokines and angiogenic factors by macrophages and MSCs. Deposition of dexamethasone-releasing coatings on polymer/Mg composites appears to be a promising approach to delay composite degradation at the early stage of implantation and may be useful to attenuate inflammation and adverse foreign body reactions.