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1.
Colloids Surf B Biointerfaces ; 207: 111989, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34303114

RESUMO

Hydrogels based on short peptide molecules are interesting biomaterials with wide present and prospective use in biotechnologies. A well-known possible drawback of these materials can be their limited mechanical performance. In order to overcome this problem, we prepared Fmoc-Phe3self-assembling peptides by a biocatalytic approach, and we reinforced the hydrogel with graphene oxide nanosheets. The formulation here proposed confers to the hydrogel additional physicochemical properties without hampering peptide self-assembly. We investigated in depth the effect of nanocarbon morphology on hydrogel properties (i.e. morphology, viscoelastic properties, stiffness, resistance to an applied stress). In view of further developments towards possible clinical applications, we have preliminarily tested the biocompatibility of the composites. Our results showed that the innovative hydrogel composite formulation based on FmocPhe3 and GO is a biomaterial with improved mechanical properties that appears suitable for the development of biotechnological applications.


Assuntos
Grafite , Hidrogéis , Peptídeos , Estudos Prospectivos
2.
Materials (Basel) ; 14(9)2021 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-33922799

RESUMO

Maxillary sinus augmentation is often necessary prior to implantology procedure, in particular in cases of atrophic posterior maxilla. In this context, bone substitute biomaterials made of biphasic calcium phosphates, produced by three-dimensional additive manufacturing were shown to be highly biocompatible with an efficient osteoconductivity, especially when combined with cell-based tissue engineering. Thus, in the present research, osteoinduction and osteoconduction properties of biphasic calcium-phosphate constructs made by direct rapid prototyping and engineered with ovine-derived amniotic epithelial cells or amniotic fluid cells were evaluated. More in details, this preclinical study was performed using adult sheep targeted to receive scaffold alone (CTR), oAFSMC, or oAEC engineered constructs. The grafted sinuses were explanted at 90 days and a cross-linked experimental approach based on Synchrotron Radiation microCT and histology analysis was performed on the complete set of samples. The study, performed taking into account the distance from native surrounding bone, demonstrated that no significant differences occurred in bone regeneration between oAEC-, oAFMSC-cultured, and Ctr samples and that there was a predominant action of the osteoconduction versus the stem cells osteo-induction. Indeed, it was proven that the newly formed bone amount and distribution decreased from the side of contact scaffold/native bone toward the bulk of the scaffold itself, with almost constant values of morphometric descriptors in volumes more than 1 mm from the border.

3.
Stem Cells Int ; 2021: 6632052, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33688354

RESUMO

Musculoskeletal regenerative medicine is mainly based on the use of cell therapy to heal damaged tissues such as bone, cartilage, and tendons. Throughout the years, different cell types have been employed for the treatment of musculoskeletal diseases, in particular, mesenchymal stem cells (MSCs) derived from bone marrow (BMSCs) and adipose tissue (ADSCs). Though the results of these literature studies have been encouraging, there are some limitations, especially on long-term results. Recently, some interest has shifted towards new cell types such as the stromal vascular fraction (SVF) and amniotic endothelial cells (AECs). The aim of the present literature review is to evaluate preclinical and clinical studies that used SVF and AECs for musculoskeletal tissue regeneration. Forty-eight preclinical and clinical studies, performed in the last 10 years, were identified. Both SVF and AECs, injected or implanted with or without scaffolds, were shown to be valid alternatives, and in some ways superior, to ADSCs and BMSCs, being able to differentiate towards osteogenic, chondrogenic, and tenogenic lineages, and to promote cell and tissue regenerative potential. The use of SVF and AECs could represent a new regenerative treatment in several musculoskeletal pathologies, solving the problem of cell expansion in vitro.

4.
Int Orthop ; 45(2): 427-435, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-32661637

RESUMO

PURPOSE: Aiming to prevent cartilage damage during early osteoarthritis (OA), the therapeutic challenge is to restore and maintain the physiological and functional properties of such a tissue with minimally invasive therapeutic strategies. METHODS: Accordingly, an in vivo model of early OA in sheep was here treated through three different cell therapies (culture expanded ADSCs, SVF, and culture expanded AECs) thus to preserve the joint surface from the progression of the pathology. Three months after the treatment injections, their performance was assessed through mechanical automated mapping (Young's modulus and cartilage thickness), gross evaluation of articular surfaces, and biochemical analysis of the synovial fluid. RESULTS: No severe degeneration was observed after three months from OA induction. Cartilage mechanical properties were crucial to identify early degeneration. All the treatments improved the macroscopic cartilage surface aspect and reduced pro-inflammatory cytokines in the synovial fluid. Among the three treatments, SVF highlighted the best performance while ADSCs the worst. CONCLUSION: Despite that the evaluated experimental time is an early follow-up and, thus, longer trial is mandatory to properly assess treatments effectiveness, the proposed multidisciplinary approach allowed to obtain preliminary, but also crucial, results concerning the reduction in OA signs on cartilage properties, in osteophyte development and in all the inflammatory markers.


Assuntos
Cartilagem Articular , Osteoartrite do Joelho , Osteoartrite , Animais , Fenômenos Biomecânicos , Modelos Animais de Doenças , Articulação do Joelho/cirurgia , Osteoartrite/terapia , Osteoartrite do Joelho/terapia , Ovinos
5.
Cells ; 8(10)2019 09 20.
Artigo em Inglês | MEDLINE | ID: mdl-31547126

RESUMO

Osteoarthritis is a degenerative disease that strongly correlates with age and promotes the breakdown of joint cartilage and subchondral bone. There has been a surge of interest in developing cell-based therapies, focused particularly on the use of mesenchymal stromal cells (MSCs) isolated from adult tissues. It seems that MSCs derived from synovial joint tissues exhibit superior chondrogenic ability, but their unclear distribution and low frequency actually limit their clinical application. To date, the influence of aging on synovial joint derived MSCs' biological characteristics and differentiation abilities remains unknown, and a full understanding of the mechanisms involved in cellular aging is lacking. The aim of this study was therefore to investigate the presence of age-related alterations in synovial fluid MSCs and their influence on the potential ability of MSCs to differentiate toward chondrogenic phenotypes. Synovial fluid MSCs, isolated from healthy equine donors from 3 to 40 years old, were cultured in vitro and stimulated towards chondrogenic differentiation for up to 21 days. An equine model was chosen due to the high degree of similarity of the anatomy of the knee joint to the human knee joint and as spontaneous disorders develop that are clinically relevant to similar human disorders. The results showed a reduction in cell proliferation correlated with age and the presence of age-related tetraploid cells. Ultrastructural analysis demonstrated the presence of morphological features correlated with aging such as endoplasmic reticulum stress, autophagy, and mitophagy. Alcian blue assay and real-time PCR data showed a reduction of efficiency in the chondrogenic differentiation of aged synovial fluid MSCs compared to young MSCs. All these data highlighted the influence of aging on MSCs' characteristics and ability to differentiate towards chondrogenic differentiation and emphasize the importance of considering age-related alterations of MSCs in clinical applications.


Assuntos
Envelhecimento/patologia , Diferenciação Celular , Condrócitos/fisiologia , Células-Tronco Mesenquimais/fisiologia , Osteoartrite/patologia , Envelhecimento/fisiologia , Animais , Proliferação de Células , Células Cultivadas , Senescência Celular/fisiologia , Condrócitos/patologia , Condrogênese/fisiologia , Modelos Animais de Doenças , Feminino , Cavalos , Humanos , Masculino , Células-Tronco Mesenquimais/patologia , Osteoartrite/fisiopatologia , Líquido Sinovial/citologia
6.
J Craniofac Surg ; 30(4): 1073-1077, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30676452

RESUMO

The poly D,L-lactide-co-glycolic acid (PLGA) is a copolymer used in many therapeutic devices for its high rates of biodegradability and biocompatibility. The principal aim of the research was to evaluate the new bone formation, after 16 (T1) and 28 weeks (T2), in sheep maxillary sinus lift in vivo model using PLGA.Computerized tomography analysis, X-ray microanalysis, and scanning electron microscope analysis of secondary electrons (SE) and the backscattered electrons (BSE) of the samples were detected.After 28 weeks, the computed tomography analysis showed a 22% increase of UH density in the grafting areas. The X-ray microanalysis of the samples showed calcium and phosphorus increase at T1 and T2 follow-up period and the carbon and oxygen concentration decrease. The SE evaluation showed a rapid superficial resorption of the biomaterials at T1 and a completely bone reorganization of biomaterial at T2. The BSE analysis confirmed the SE data and showed the direct and intimate contact between bone and PLGA with a higher calcification in T2 compared to T1.Certainly, still other experiments and a larger number of samples will be necessary to be analyzed to determine the behavior of the PLGA in the bone regeneration; however, the PLGA used in maxillary sinus lift animal model, seem to promote new bone formation that continues increase at 28 weeks after grafting.


Assuntos
Regeneração Óssea/efeitos dos fármacos , Glicolatos/farmacologia , Seio Maxilar/cirurgia , Animais , Materiais Biocompatíveis , Regeneração Óssea/fisiologia , Calcificação Fisiológica , Implantes Dentários , Seio Maxilar/diagnóstico por imagem , Seio Maxilar/fisiologia , Modelos Animais , Ovinos , Levantamento do Assoalho do Seio Maxilar , Tomografia Computadorizada por Raios X
7.
Sports Med Arthrosc Rev ; 26(2): 70-74, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29722767

RESUMO

This is a study of amniotic epithelial cells, which form the innermost layer of the amniotic membrane. These cells can be easily isolated and display peculiar and unique properties, such as plasticity and differentiation potential toward the 3 germinal layers, that may aid regeneration and/or repair of damaged or diseased tissues and organs. A robust literature based on in vitro, experimental, and clinical studies in large animals demonstrates that these cells can enhance the regeneration of tendons, bone, and articular cartilage. On the basis of these considerations, allotransplantation of human amniotic epithelial cells could be proposed for clinical trials in human orthopedic conditions.


Assuntos
Âmnio/citologia , Terapia Baseada em Transplante de Células e Tecidos/tendências , Células Epiteliais/citologia , Células-Tronco/citologia , Animais , Doenças Ósseas/terapia , Doenças das Cartilagens/terapia , Humanos , Traumatismos dos Tendões/terapia
8.
J Tissue Eng Regen Med ; 12(3): e1594-e1608, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29024514

RESUMO

Cell-based therapy holds great promise for tendon disorders, a widespread debilitating musculoskeletal condition. Even if the cell line remains to be defined, preliminary evidences have proven that amniotic-derived cells possess in vitro and in vivo a great tenogenic potential. This study investigated the efficacy of transplanted human amniotic epithelial cells (hAECs) by testing their early regenerative properties and mechanisms involved on a validated ovine Achilles tendon partial defect performed on 29 animals. The injured tendons treated with hAECs recovered rapidly, in 28 days, structural and biomechanical properties undertaking a programmed tissue regeneration, differently from the spontaneous healing tissues. hAECs remained viable within the host tendons establishing with the endogenous progenitor cells an active dialogue. Through the secretion of modulatory factors, hAECs inhibited the inflammatory cells infiltration, activated the M2 macrophage subpopulation early recruitment, and accelerated blood vessel as well as extracellular matrix remodelling. In parallel, some in situ differentiated hAECs displayed a tenocytelike phenotype. Both paracrine and direct hAECs stimulatory effects were confirmed analysing their genome profile before and after transplantation. The 49 human up-regulated transcripts recorded in transplanted hAECs belonged to tendon lineage differentiation (epithelial-mesenchymal transition, connective specific matrix components, and skeleton or muscle system development-related transcripts), as well as the in situ activation of paracrine signalling involved in inflammatory and immunomodulatory response. Altogether, these evidences support the hypothesis that hAECs are a practicable and efficient strategy for the acute treatment of tendinopathy, reinforcing the idea of a concrete use of amniotic epithelial cells towards the clinical practice.


Assuntos
Tendão do Calcâneo/patologia , Âmnio/citologia , Células Epiteliais/transplante , Regeneração , Tendão do Calcâneo/irrigação sanguínea , Tendão do Calcâneo/fisiopatologia , Animais , Fenômenos Biomecânicos , Diferenciação Celular , Sobrevivência Celular , Células Epiteliais/citologia , Matriz Extracelular/metabolismo , Regulação da Expressão Gênica , Humanos , Macrófagos/metabolismo , Neovascularização Fisiológica , Fenótipo , Ovinos , Tenócitos/patologia , Transplante Heterólogo , Remodelação Vascular , Cicatrização
9.
Sci Rep ; 7(1): 3761, 2017 06 19.
Artigo em Inglês | MEDLINE | ID: mdl-28630448

RESUMO

The in vitro expansion is detrimental to therapeutic applications of amniotic epithelial cells (AEC), an emerging source of fetal stem cells. This study provides molecular evidences of progesterone (P4) role in preventing epithelial-mesenchymal transition (EMT) in ovine AEC (oAEC). oAEC amplified under standard conditions spontaneously acquired mesenchymal properties through the up-regulation of EMT-transcription factors. P4 supplementation prevented phenotype shift by inhibiting the EMT-inducing mechanism such as the autocrine production of TGF-ß and the activation of intracellular-related signaling. The effect of P4 still persisted for one passage after steroid removal from culture as well as steroid supplementation promptly reversed mesenchymal phenotype in oAEC which have experienced EMT during amplification. Furthermore, P4 promoted an acute up-regulation of pluripotent genes whereas enhanced basal and LPS-induced oAEC anti-inflammatory response with an increase in anti-inflammatory and a decrease in pro-inflammatory cytokines expression. Altogether, these results indicate that P4 supplementation is crucial to preserve epithelial phenotype and to enhance biological properties in expanded oAEC. Therefore, an innovative cultural approach is proposed in order to improve therapeutic potential of this promising source of epithelial stem cells.


Assuntos
Âmnio/imunologia , Células Epiteliais/imunologia , Transição Epitelial-Mesenquimal/imunologia , Imunomodulação , Progesterona/imunologia , Animais , Feminino , Ovinos
10.
Stem Cells Int ; 2017: 5651287, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28469672

RESUMO

Background. Neural crest-derived mesenchymal stem cells (MSCs) from human oral tissues possess immunomodulatory and regenerative properties and are emerging as a potential therapeutic tool to treat diverse diseases, such as multiple sclerosis, myocardial infarction, and connective tissue damages. In addition to cell-surface antigens, dental MSCs express embryonic stem cell markers as neural crest cells originate from the ectoderm layer. In vitro passages may eventually modify these embryonic marker expressions and other stemness properties, including proliferation. In the present study, we have investigated the expression of proteins involved in cell proliferation/senescence and embryonic stem cell markers during early (passage 2) and late passages (passage 15) in MSCs obtained from human gingiva, periodontal, and dental pulp tissues. Methods. Cell proliferation assay, beta galactosidase staining, immunocytochemistry, and real-time PCR techniques were applied. Results. Cell proliferation assay showed no difference between early and late passages while senescence markers p16 and p21 were considerably increased in late passage. Embryonic stem cell markers including SKIL, MEIS1, and JARID2 were differentially modulated between P2 and P15 cells. Discussion. Our results suggest that the presence of embryonic and proliferation markers even in late passage may potentially endorse the application of dental-derived MSCs in stem cell therapy-based clinical trials.

11.
J Periodontal Implant Sci ; 46(5): 303-319, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27800213

RESUMO

PURPOSE: The purpose of this animal study was to perform a 3-dimensional micro-computed tomography (micro-CT) analysis in order to investigate the influence of root surface distance to the alveolar bone and the periodontal ligament on periodontal wound healing after a guided tissue regeneration (GTR) procedure. METHODS: Three adult Sus scrofa domesticus specimens were used. The study sample included 6 teeth, corresponding to 2 third mandibular incisors from each animal. After coronectomy, a circumferential bone defect was created in each tooth by means of calibrated piezoelectric inserts. The experimental defects had depths of 3 mm, 5 mm, 7 mm, 9 mm, and 11 mm, with a constant width of 2 mm. One tooth with no defect was used as a control. The defects were covered with a bioresorbable membrane and protected with a flap. After 6 months, the animals were euthanised and tissue blocks were harvested and preserved for micro-CT analysis. RESULTS: New alveolar bone was consistently present in all experimental defects. Signs of root resorption were observed in all samples, with the extent of resorption directly correlated to the vertical extent of the defect; the medial third of the root was the most commonly affected area. Signs of ankylosis were recorded in the defects that were 3 mm and 7 mm in depth. Density and other indicators of bone quality decreased with increasing defect depth. CONCLUSIONS: After a GTR procedure, the periodontal ligament and the alveolar bone appeared to compete in periodontal wound healing. Moreover, the observed decrease in bone quality indicators suggests that intrabony defects beyond a critical size cannot be regenerated. This finding may be relevant for the clinical application of periodontal regeneration, since it implies that GTR has a dimensional limit.

12.
Res Vet Sci ; 105: 92-102, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27033915

RESUMO

Recently, we have demonstrated that ovine amniotic epithelial cells (oAECs) allotransplanted into experimentally induced tendon lesions are able to stimulate tissue regeneration also by reducing leukocyte infiltration. Amongst leukocytes, macrophages (Mφ) M1 and M2 phenotype cells are known to mediate inflammatory and repairing processes, respectively. In this research it was investigated if, during tendon regeneration induced by AECs allotransplantation, M1Mφ and M2Mφ phenotype cells are recruited and differently distributed within the lesion site. Ovine AECs treated and untreated (Ctr) tendons were explanted at 7, 14, and 28 days and tissue microarchitecture was analyzed together with the distribution and quantification of leukocytes (CD45 positive), Mφ (CD68 pan positive), and M1Mφ (CD86, and IL12b) and M2Mφ (CD206, YM1 and IL10) phenotype related markers. In oAEC transplanted tendons CD45 and CD68 positive cells were always reduced in the lesion site. At day 14, oAEC treated tendons began to recover their microarchitecture, contextually a reduction of M1Mφ markers, mainly distributed close to oAECs, and an increase of M2Mφ markers was evidenced. CD206 positive cells were distributed near the regenerating areas. At day 28 oAECs treated tendons acquired a healthy-like structure with a reduction of M2Mφ. Differently, Ctr tendons maintained a disorganized morphology throughout the experimental time and constantly showed high values of M1Mφ markers. These findings indicate that M2Mφ recruitment could be correlated to tendon regeneration induced by oAECs allotransplantation. Moreover, these results demonstrate oAECs immunomodulatory role also in vivo and support novel insights into their allogeneic use underlying the resolution of tendon fibrosis.


Assuntos
Âmnio/citologia , Transplante de Células/veterinária , Macrófagos/fisiologia , Regeneração , Traumatismos dos Tendões/veterinária , Tendões/fisiologia , Animais , Transplante de Células/métodos , Células Epiteliais/citologia , Fenótipo , Ovinos , Traumatismos dos Tendões/terapia
13.
J Craniofac Surg ; 26(3): 737-40, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25974782

RESUMO

Sinus augmentation is a routine surgical procedure in dentistry. At present, various animal models are available for the research purpose on this topic. In particular, for the first time, we have performed a morphological study on sheep sinus, using cone beam computed tomography (CBCT), to precisely define the anatomy of the ovine sinus. Then, we compared the sheep and human sinus morphological parameters, in order to uniform the research approach to the sinus augmentation procedures and to standardize this experimental model. Six fresh heads of adult female sheep were studied with CBCT and histologic examination to determine the dimensions and the organization of the ovine maxillary sinus. The comparison of the dimensional values between man and sheep shows evident differences between the two species; CBCT offers detailed information for studying normal maxillary sinus. Human and sheep maxillary sinus show anatomical differences that must be taken into account in experimental procedures.


Assuntos
Modelos Animais , Levantamento do Assoalho do Seio Maxilar/métodos , Pesquisa Translacional Biomédica , Adulto , Animais , Tomografia Computadorizada de Feixe Cônico/métodos , Feminino , Humanos , Seio Maxilar/diagnóstico por imagem , Seio Maxilar/cirurgia , Ovinos
14.
Muscles Ligaments Tendons J ; 5(1): 5-11, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25878980

RESUMO

BACKGROUND: amnion-derived stem cells are considered a promising alternative source for tendon tissue regeneration. PURPOSE: aims of this paper were to illustrate the ultrasound and histological outcomes following the treatment of acute and chronic superficial digital flexor tendon spontaneous lesions in horses with ovine amniotic epithelial cells xenotransplantation. METHODS: six adult horses suffering from unilateral acute (4 cases) and chronic (2 cases) tendinopathy (clinical and ultrasound diagnosis) were enrolled. At baseline, ovine amniotic epithelial cells were grafted, in sterile conditions and under ultra-sound control, into the most damaged area. Ultra-sound controls were performed at 30, 60, 90, 120, 150 and 180 days after cells implantation; after horse euthanasia (180 days) tendon samples were collected and submitted to histological examination (cellularity, extracellular matrix fiber organization, blood vessels). RESULTS: at baseline, in the acute cases, the ultra-sound exam showed a focal, dis-homogeneous, hypo-echoic area into the superficial digital flexor tendon, with loss of the normal fibrillar pattern, while in the chronic cases the damaged tendon area appeared thickened and completely hyper-echoic. At the final follow-up tendon echotexture was more regular, the cross-sectional area similar to the contra-lateral limb, and the collagen fibers were oriented in parallel to the longitudinal axis of the tendon both in the acute and chronic cases, suggesting a positive healing response. These findings were supported by the histological analyses which showed an almost complete restoration of normal tendon architecture with an optimal alignment of tendon fibers. CONCLUSIONS: the present pilot study supports the hypothesis that amniotic epithelial cells are provided of an excellent healing potential and shows a very good correlation between the ultrasound findings and the histologic features.

15.
J Oral Implantol ; 41(3): 240-50, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23829685

RESUMO

In this study, the in vivo behavior of a custom-made three-dimensional (3D) synthetic bone substitute was evaluated when used as scaffold for sinus augmentation procedures in an animal model. The scaffold was a calcium phosphate ceramic fabricated by the direct rapid prototyping technique, dispense-plotting. The geometrical and chemical properties of the scaffold were first analyzed through light and electron scanning microscopes, helium picnometer, and semi-quantitative X-ray diffraction measurements. Then, 6 sheep underwent monolateral sinus augmentation with the fabricated scaffolds. The animals were euthanized after healing periods of 45 and 90 days, and block sections including the grafted area were obtained. Bone samples were subjected to micro computerized tomography, morphological and histomorphometric analyses. A complete integration of the scaffold was reported, with abundant deposition of newly formed bone tissue within the biomaterial pores. Moreover, initial foci of bone remodeling were mainly localized at the periphery of the implanted area after 45 days, while continuous bridges of mature lamellar bone were recorded in 90-day specimens. This evidence supports the hypothesis that bone regeneration proceeds from the periphery to the center of the sinus cavity. These results showed how a technique allowing control of porosity, pore design, and external shape of a ceramic bone substitute may be valuable for producing synthetic bone grafts with good clinical performances.


Assuntos
Regeneração Óssea , Substitutos Ósseos , Transplante Ósseo , Animais , Materiais Biocompatíveis , Fosfatos de Cálcio , Cerâmica , Ovinos
16.
Vet Ital ; 50(4): 249-57, 2014 Dec 29.
Artigo em Inglês | MEDLINE | ID: mdl-25546062

RESUMO

Biophysical stimulations with electric and electromagnetic fields have been demonstrated to accelerate the bone-healing rate. This study has been designed to investigate the effects of electricity directly connected with the central pins of an external fixator in an experimental osteotomy model in sheep. Thirty mg/kg of tetracycline chloride were administered on the 30th and on the 45th day after surgery for histomorphometric studies. Plain radiographs were obtained in standard projections every 15 days after surgery and were analyzed with a software program (Corel Photo-Paint Pro X2, Corel Corporation, Ottawa, Canada). The specimens obtained after 60 days were examined with histological analysis. The results show that biophysical treatment with alternating electricity in combination with external fixator enhances new-bone formation. The translational value of this study, due to the similarities between ovine and human species, suggests that this treatment could be useful in speeding the bone-healing rate both in animals and humans.


Assuntos
Estimulação Elétrica , Fixação de Fratura , Consolidação da Fratura , Fraturas Ósseas/terapia , Animais , Terapia Combinada , Modelos Animais de Doenças , Fixadores Externos , Ovinos
17.
PLoS One ; 8(5): e63256, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23696804

RESUMO

BACKGROUND: Evidence has been provided that a cell-based therapy combined with the use of bioactive materials may significantly improve bone regeneration prior to dental implant, although the identification of an ideal source of progenitor/stem cells remains to be determined. AIM: In the present research, the bone regenerative property of an emerging source of progenitor cells, the amniotic epithelial cells (AEC), loaded on a calcium-phosphate synthetic bone substitute, made by direct rapid prototyping (rPT) technique, was evaluated in an animal study. MATERIAL AND METHODS: Two blocks of synthetic bone substitute (∼0.14 cm(3)), alone or engineered with 1×10(6) ovine AEC (oAEC), were grafted bilaterally into maxillary sinuses of six adult sheep, an animal model chosen for its high translational value in dentistry. The sheep were then randomly divided into two groups and sacrificed at 45 and 90 days post implantation (p.i.). Tissue regeneration was evaluated in the sinus explants by micro-computer tomography (micro-CT), morphological, morphometric and biochemical analyses. RESULTS AND CONCLUSIONS: The obtained data suggest that scaffold integration and bone deposition are positively influenced by allotransplantated oAEC. Sinus explants derived from sheep grafted with oAEC engineered scaffolds displayed a reduced fibrotic reaction, a limited inflammatory response and an accelerated process of angiogenesis. In addition, the presence of oAEC significantly stimulated osteogenesis either by enhancing bone deposition or making more extent the foci of bone nucleation. Besides the modulatory role played by oAEC in the crucial events successfully guiding tissue regeneration (angiogenesis, vascular endothelial growth factor expression and inflammation), data provided herein show that oAEC were also able to directly participate in the process of bone deposition, as suggested by the presence of oAEC entrapped within the newly deposited osteoid matrix and by their ability to switch-on the expression of a specific bone-related protein (osteocalcin, OCN) when transplanted into host tissues.


Assuntos
Líquido Amniótico/citologia , Regeneração Óssea , Substitutos Ósseos , Células Epiteliais/transplante , Seio Maxilar/cirurgia , Animais , Células Cultivadas , Células Epiteliais/metabolismo , Expressão Gênica , Maxila/fisiologia , Maxila/cirurgia , Seio Maxilar/irrigação sanguínea , Carneiro Doméstico , Engenharia Tecidual , Alicerces Teciduais , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , Microtomografia por Raio-X
18.
Theriogenology ; 80(2): 120-4, 2013 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-23611407

RESUMO

The donkey reproductive physiology is still partially known despite the increasing risk of extinction involving several breeds. The present study was designed to describe the postpartum (PP) reproductive performance of an Italian endangered breed: the Martina Franca donkey. To this aim, 52 jennies were monitored to define the foal-heat (FH) and the first and second PP estrus episodes (1st PPe and 2nd PPe). The data indicate that jennies spontaneously recovered reproduction in approximately 10 days after delivery. Then heats occur with a regular interval of approximately 23 days. Estrus length was 1 week in FH and the 2nd PPe and significantly shorter in the 1st PPe. Estrus-ovulation, and delivery-ovulation interval and follicle growth were similar in all animals tested. Pregnancy rate (PR) was lower when natural mating occurred during the FH and 2nd PPe (approximately 60%) than during the 1st PPe (approximately 70%; P < 0.01). In addition, the higher PR (>80%; P < 0.01) was recorded in jennies when the FH occurred after the first week PP and it dropped (<50%) in early FH animals. The PR was also affected by the season and by age: it significantly declined during the autumn-winter season and in subjects older than the sixth year of age. For the first time, the reproductive performance of PP donkeys were defined on a large number of Martina Franca jennies thus offering useful information to improve farm management with an immediate benefit to increase livestock production. This aspect of management improvement might be particularly important if applied to an endangered breed such as Martina Franca donkeys.


Assuntos
Espécies em Perigo de Extinção , Equidae/fisiologia , Período Pós-Parto/fisiologia , Prenhez , Reprodução/fisiologia , Animais , Cruzamento , Feminino , Itália , Ovulação/fisiologia , Parto/fisiologia , Gravidez , Fatores de Tempo
19.
Clin Oral Investig ; 17(7): 1661-75, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23064983

RESUMO

OBJECTIVES: The present research has been performed to evaluate whether a commercial magnesium-enriched hydroxyapatite (MgHA)/collagen-based scaffold engineered with ovine amniotic fluid mesenchymal cells (oAFMC) could improve bone regeneration process in vivo. MATERIALS AND METHODS: Bilateral sinus augmentation was performed on eight adult sheep in order to compare the tissue regeneration process at 45 and 90 days after implantation of the oAFMC-engineered scaffold (Test Group) or of the scaffold alone (Ctr Group). The process of tissue remodeling was analyzed through histological, immunohistochemical, and morphometric analyses by calculating the proliferation index (PI) of oAFMC loaded on the scaffold, the total vascular area (VA), and vascular endothelial growth factor (VEGF) expression levels within the grafted area. RESULTS: MgHA/collagen-based scaffold showed high biocompatibility preserving the survival of oAFMC for 90 days in grafted sinuses. The use of oAFMC increased bone deposition and stimulated a more rapid angiogenic reaction, thus probably supporting the higher cell PI recorded in cell-treated sinuses. A significantly higher VEGF expression (Test vs. Ctr Group; p = 0.0004) and a larger total VA (p = 0.0006) were detected in the Test Group at 45 days after surgery. The PI was significantly higher (p = 0.027) at 45 days and became significantly lower at 90 days (p = 0.0007) in the Test Group sinuses, while the PI recorded in the Ctr Group continued to increase resulting to a significantly higher PI at day 90 (CTR day 45 vs. CTR day 90; p = 0.022). CONCLUSIONS: The osteoinductive effect of a biomimetic commercial scaffold may be significantly improved by the presence of oAFMC. CLINICAL RELEVANCE: The amniotic fluid mesenchymal cell (AFMC) may represent a novel, largely and easily accessible source of mesenchymal stem cells to develop cell-based therapy for maxillofacial surgery.


Assuntos
Líquido Amniótico/citologia , Transplante de Células-Tronco Mesenquimais , Levantamento do Assoalho do Seio Maxilar/métodos , Engenharia Tecidual/métodos , Aloenxertos , Animais , Materiais Biocompatíveis , Regeneração Óssea , Proliferação de Células , Durapatita/farmacologia , Citometria de Fluxo , Imuno-Histoquímica , Magnésio/farmacologia , Modelos Animais , Osteogênese , Carneiro Doméstico , Alicerces Teciduais , Fator A de Crescimento do Endotélio Vascular/metabolismo
20.
Muscles Ligaments Tendons J ; 2(3): 187-92, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23738296

RESUMO

The ideal strategy for tendon healing has not been identified to date. Recently, the use of stem cells based therapy has been proposed, due to their ability to proliferate and to differentiate towards specific connective tissues lineages. Embryonic stem cells should be considered the ideal cell source for regenerative therapies, but ethical factors limit their use in humans. Mesenchymal stem cells are more easily available and can be obtained by different sources. Amnion derived stem cells can differentiate towards all three germ layers, and can be used for allogeneic transplantation and stored thanks to cryopreservation. In veterinary medicine, stem cells have been used with encouraging results for the treatment of the Superficial Digital Flexor tendinopathy in the horses. Considering that Superficial Digital Flexor tendinopathy is similar for pathogenesis and histopathology to Achilles tendinopathy in man, this experience can provide supportive data to encourage the use of regenerative therapy in humans.

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