1,721,036 research outputs found
FROM IN VITRO STUDIES TO A LARGE ANIMAL MODEL: A MULTISTEP DISSECTION ON THE FUTURE ROLE OF ADIPOSE-DERIVED STEM CELLS FOR MUSCULOSKELETAL TISSUE ENGINEERING.
Full text linkTissue engineering is an emerging interdisciplinary field, born with the purpose to provide an alternative solution for the regeneration of lesioned or lost tissues, combining cells, biocompatible scaffolds and bioactive factors. The cells for this approach should be non-immunoreactive and non-tumorigenic. Moreover, they should be available in large amount and possess, or be able to acquire, a specific protein expression pattern similar to that of the damaged tissue and/or act as a pool of trophic factors for resident cells. All these reasons, make mesenchymal stem cells (MSCs) good candidates for applications in regenerative medicine. Although bone marrow is still the most common source of MSCs, these cells could be harvested from all vascularised tissues, and, interestingly, from tissues that are normally discarded, such as fat, placenta or umbilical cord. One of the most convenient source of MSCs, is unequivocally, the adipose tissue due to the easily accessible anatomical location and the abundance of subcutaneous adipose tissue. Adipose-derived stem cells (ASCs) are similar to MSCs isolated from bone marrow in morphology, immunophenotype, and differentiation ability, and own interesting features such as immunoregolatory and anti-inflammatory properties.
In the recent years, many strategies for the cure of musculoskeletal tissues critical lesions, mainly in orthopaedic, oral and maxillo-facial surgery, have been under investigations. In this contest, the regeneration of structures including different tissues, such as the periodontium and the osteochondral unit, are particularly challenging.
Periodontal regeneration is especially demanding, as it requires regeneration of three quite diverse and unique tissues such as the alveolar bone, the periodontal ligament and the cementum, that have to interface with each other to restore their complex structure. Since the promising results obtained with ASCs in preclinical studies of periodontal diseases arouse the curiosity of maxillofacial and dental surgeons, we decided to identify a novel source of ASCs, i.e, the buccal fat pad, convenient for these specialists. For this purpose, we studied human adipose derived-stem cells from buccal fat pad (BFP-ASCs), comparing them with cells from the subcutaneous adipose tissue (SC-ASCs) of the same donor (n=2). In parallel, considering the need for preclinical studies in which the effect of allogenic cells should be tested, and swine as an accepted animal model in tissue engineering applications, we also characterized porcine cells (n=6). With preclinical and clinical application prospective, we also investigated ASC interactions with oral tissues, natural and synthetic scaffolds and Amelogenin, an oral bioactive molecule. First of all, we showed that it is feasible to isolate ASCs even starting from very limited amounts of tissue (0,5 ml) and that the cellular yield is influenced by species, but not by the site of harvesting (1.1x105±1.4x104 human BFP-ASCs/ml and 1.15x105±7.1x103 human SC-ASCs/ml; 3.0x104±9.3x103 porcine BFP-ASCs/ml and 5.5x104±3.3x104 porcine SC-ASCs/ml). Despite the lower yield, the pASCs great proliferation rate allows to obtain high number of cells (potentially, 108 - 109) after few (3, 4) passages in culture. After the isolation, a great amount of cells deriving from all the tissues, adhered to cell culture plates showing the MSC fibroblast like morphology, with only mild shape differences constituted by the higher elongation and dimension of human SC-ASCs. Moreover, all the cells are easily expandable and showed good clonogenic ability at early passages. Cells of the same species, from both the harvesting site, displayed the same surface markers profile, that, in particular for human ASCs, was the typical one of hMSC (CD90+, CD105+, CD73+, CD14-, CD31-, and CD34-). Human and porcine BFP-ASCs, as SC-ASCs, are multipotent; indeed, when induced towards osteogenic and adipogenic lineages, they up-regulated significantly ALP activity, collagen and calcified extracellular matrix deposition and lipid vacuoles productions, respectively, already after 14 days of differentiation in vitro.
Next, since cell/scaffold interaction is fundamental for the outcome of a tissue engineering approach, in sight of a preclinical study, we combined porcine BFP and SC-ASCs to both clinical grade (titanium) and innovative [silicon carbide–plasma-enhanced chemical vapor deposition (SIC-PECVD)] biomaterials, and studied cell adhesion and their differentiation ability. All the cells nicely grew on both scaffolds and, when osteoinduced, significantly increased the amount of calcified ECM compared to control cells; interestingly, titanium is osteoinductive even per se on pASCs (+284% and +91 for BFP- and SC-ASCs). Considering the importance of cell interaction with tissue of the lesion site, and with materials commonly used during surgical practices, we studied human BFP- and SC-ASC adherence to several supports. SEM analysis confirmed that both cell type nicely stick on alveolar bone, periodontal ligament, collagen membrane and polyglycolic acid filaments. Finally, we found that amelogenin, the most abundant enamel matrix protein seems to be an early osteoinductive factor for BFP-hASCs, whereas this effect is not manifested for SC-hASCs.
For future cellular therapy, and since the use of FBS pose the risk of xenogenic contaminations leading to immunological complications during transplant, we tested cells growth in the presence of autologous supplements. Interestingly, both hASCs adapted rapidly to human serum, increasing their proliferation rates compared to standard culture condition, while porcine autologous or heterologous sera, did not improve pASC growth.
In conclusion, we identified a cell population derived from a tissue easily available to dentists and maxillofacial surgeons, whose multipotent features and interaction with clinical grade scaffolds make proper candidate for future uses in tissue engineering approaches of periodontal diseases.
In parallel, part of my PhD project was focused on the study of a critical osteochondral defect regeneration performed in a large animal preclinical model.
The main obstacles for clinicians in treating this defect arises from the disparity concerning anatomy, composition and, most importantly, rate of healing of the articular cartilage (AC) and the subchondral bone. The key points of our study are the use of an innovative hydrogel of oligo(polyethylene glycol)fumarate (OPF) to fill the osteochondral defect, and of either porcine, or human ASCs, to create bioconstructs to be implanted in non-immuno-compromised minipigs. In particular, four critical osteochondral defects (diameter 9mm, depth 8mm) were created in the peripheral part of the trochlea of seven animals (defect n=28), and then treated with the different pre-made constructs. Untreated defects and defects filled by just scaffold were included as controls.
No side-effects have been observed during the six-moths follow-up. At the end of this period, animals were sacrificed and knees explanted. Gross appearance analyses showed quite satisfactory filling of all the lesions, with the exception of one animal, whose joint appeared infected and not healed.
MRI analyses revealed that in all the scaffold treated groups an overall improvement of the tissue quality at the osteochondral lesion site, was induced. More accurate evaluations (histological and immunohistochemistry analyses) revealed that some important tissue features were significantly improved by the association of OPF and ASCs. Indeed, regarding the subchondral bone, in all the OPF+ASCs groups, a mature bone appeared, with higher deposition of collagen type I compared to untreated or unseeded OPF groups. Moreover, the use of ASCs associated to scaffolds induced an improvement in newly formed cartilage features such as collagen type II deposition, and histological scores associated to these samples indicated a significant increase in matrix staining, tissue morphology and formation of tidemark, together with a reduction in vascularisation (a positive aspect in cartilage) compared to unseeded scaffolds.
However, the histology indicated that in all the samples cartilage regeneration was still immature, most likely due to the limited time of follow up and/or the insufficient stimuli for cartilage complete regeneration. Despite this, biomechanical tests revealed that the neo-cartilage found in the cell-loaded scaffold groups possessed poroelastic behaviour, as well as indentation modulus and creep curves comparable to native cartilage. This important result suggest that the ASC presence at the lesion site, is able to enhance newly formed cartilage functionality.
In conclusion, this in vivo study provide the evidence that both porcine and human adipose-derived stem cells associated to OPF hydrogel improve osteochondral defect regeneration, even though, at the moment, we are not able to define if the implanted ASCs are responsible per se of the new tissue formation or if they help spontaneous regeneration process by paracrine actions
Rabbit Adipose-derived Stem Cells and tibia repair
No full textAdipose-derived stem cells (ASCs) may represent, alone or in combination with different scaffolds, a novel and efficient approach for bone regeneration. Here, we describe how autologous rabbit ASCs (rbASCs) isolated from interscapular adipose tissue, expanded and characterized in vitro, are used to regenerate a full-thickness bone defect in the tibial crest of New Zealand rabbits.
The animals have been divided in 4 groups: one group where the lesions have been treated with rbASCs seeded on hydroxyapatite-disk (rbASCs-HA), one group with only rbASCs, one with just HA, and one untreated group (just defect). The follow-up was of eight weeks.
Meanwhile, rbASCs have been characterized in vitro: these progenitor cells show a homogenous high proliferation rate and a marked clonogenic ability. Moreover, rbASCs demonstrate an osteogenic potential that has been evaluated by the expression of specific bone markers such as alkaline phosphatase, collagen, osteonectin and extracellular calcified matrix deposition, both in the absence and in the presence of hydroxyapatite.
Eight weeks after surgical interventions, gross appearance, X-rays, histological analyses and biomechanical tests were performed on all the animals.
The macroscopic analyses of all the tibias show a satisfactory filling of the lesions without any significant difference in terms of stiffness. By X-rays, a good osteo-integration appears in both scaffold-treated groups; despite the fact that HA was not completely resorbed, cells-HA treated bones show a more efficient scaffold resorption than the other group. In addition, the scaffold-treated defects show a better bone formation compared to the control samples. In particular, the new bone, formed in the presence of rbASCs-HA, is more mature and similar to the native one showing an improvement in bone mechanical properties.
These results indicate that autologous ASCs-hydroxyapatite bio-construct may be a potential treatment for the regeneration of bone defects
Monitoring oral and epithelial primary cells onto dental implants
No full textOBJECTIVE: Dental implants are successfully employed to treat partial or complete edentulism and the 15 year survival rate is higher than 90%. Their long-term success depends mainly on both osseointegration (the formation of an effective interface between implant and bone) and gingival attachment through junctional epithelium. Previously, scientific interest was mostly focused on bone-implant interaction, now the importance of soft tissue sealing has emerged. Indeed, to prevent mechanical instability and infection, epithelial cells must create a tight collar around the implant, as it happens physiologically around teeth. Therefore, the early attachment of gingival cells and their spreading play a critical role for the clinical outcome of dental implants. Research in dental implant material focuses on surface roughness, defined by the index of micro-irregularities Ra, which might promote osseointegration and inhibition of soft tissue attachment to the prosthesis. Here we explored the influence of surface roughness on adhesion, proliferation/viability, morphology and spreading of primary cell cultures isolated from soft tissues (human oral keratinocytes, hOKs, gingival progenitor cells, hGPCs, and epithelial cells, hECs).
MATERIALS AND METHODS: Human primary cells were seeded on three clinical grade implants with different surface characteristics: machined (S1, Ra=0.3-0.6 μm), Ti-Unite® (S2, Ra=1.2 μm) and SLA® (S3, Ra=2 μm). Adhesion, vitality and spreading were analysed combining AlamarBlue® Cell Viability Assay with microscopic techniques (stereo- and confocal microscopy after Calcein staining).
RESULTS: Cellular adhesion on S1 and S2 implants (36±2% and 44±7% respectively) was higher than on S3 (23±6%). In addition, cell proliferation slightly improved on the machined surface (+200±12% for S1, +155% for both S2 and S3). These results were confirmed by a microscopy-based analysis of cell morphology and spreading. Then, re-used biomaterials gave inconsistent results in vitro.
CONCLUSION: Implant surface affects hOK and progenitor cell adhesion and proliferation. Appropriately designed in vitro studies may be predictive of the implant clinical outcome and we propose this strategy, combining viability assays and image-based analyses, as a reproducible and rapid first step in screening new materials for clinical purposes
BMSCs and ASCs response to 17 β-estradiol (E2)
No full textEstrogens confer skeletal protective effects, facilitating osteogenesis and suppressing bone-resorption. 17β estradiol (E2) effect on progenitor cells (MSCs) has been largely studied, nevertheless data are still controversial. Since Mesenchymal Stem Cells from bone marrow (BMSCs) and adipose tissue (ASCs) are widely used in bone-tissue engineering research, we evaluated in vitro the effect of E2 on their proliferation, viability and osteogenic ability. E2 did not influence cell viability and proliferation; in particular, MSCs treated with E2 (1-1000 nM) for 14 days behaved as the untreated ones and no modifications in morphology and proliferation rate was observed after 35 days of treatment. In order to verify a pro-osteogenic effect, MSCs were pre-treated with 10, 100 nM E2 for 7, 21, 35 days and then osteo-differentiated for 14 days. During the osteo-differentiative process E2 increased significantly BMSC ALP activity (+50%), and this marker was further induced by 7 days pre-treatment (+158% 10 nM, +185% 100 nM) and maintained by longer pre-treatments. In contrast, E2 never influenced the osteogenic ability of ASCs at the doses and time points considered. Next, since E2 seems to be involved in telomerase activation, we wanted to evaluate its gene expression and enzymatic activity in our cells. We were unable to detect it by RT-PCR, and its activity was randomly modulated. Our data suggest a difference of E2 treatment on BMSC and ASC response which could be due to the different expression of one of its receptors. We analysed ERα expression by western blot and, surprisingly, an ERα variant of about 37 KDa is expressed by both BMSCs and ASCs, whereas the classic 66 KDa isoform was faintly or even not depicted. In conclusion, E2 improved osteogenic differentiation just on BMSCs without effect on ASCs; further studies are now in progress to elucidate the ERα variant function in MSCs
Study in vitro on porcine stem cells from buccal fat pad and subcutaneous adipose tissue for periodontal and oral bone
No full textOral bone lost is an important issue in maxillo-facial and dental surgery. In our previous study we have characterized human mesenchymal stem cells from Bichat’s fat pad, a convenient tissue for dentists, and proposed them for future therapies of periodontal defects and oral bone regeneration. Considering the need for preclinical study and the swine as an accepted animal model in tissue engineering applications, here we compared the swine Adipose-derived Stem Cells (ASCs) from Buccal fat pad and from subcutaneous adipose tissue both alone and in combination with synthetic scaffolds.
5.5x104±3.3x104 and 3.0x104±9.3x103 cells/ml were isolated from interscapular subcutaneous adipose tissue (ScI) and buccal fat pad (BFP) of 6 swine. All the ASCs, with MSC fibroblast-like morphology, proliferated costantly during culture with an average doubling time (DT) of 82.9±11.5 hours and 72.5±8.2 hours for ScI- and BFP-ASCs, respectively, and showed a high clonogenic ability (+10.1±1.4% for ScI- and +8.9±1.5% for BFP-ASCs). By FACS analysis both cell populations appeared similar in size and granularity, and expressed CD90, whereas the CD14, CD45 and CD271 were not detectable. Furthermore, after osteogenic induction, all the ASCs showed an increase of either collagen and calcified extracellular matrix (ECM) production of about 87% and 118% for ScI-ASCs, and of about 254% and 116% for BFP-ASCs, compared to undifferentiated cells. Alkaline phosphatase activity (+126% and +201% for ScI- and BFP-ASCs, respectively) and osteonectin expression (+336% for ScI- and +306% for BFP-ASCs) were also upregulated, confirming their capacity to osteo-differentiate. When induced, both cell types were also able to differentiate towards the adipogenic and chondrogenic lineages, as revealed by lipid vacuoles production and GAGs deposition. Additionally, the osteoinduction of cells in the presence of titanium disks and silicon carbide–plasma-enhanced chemical vapor deposition (SIC-PECVD) fragments, significantly increased the amount of calcified ECM compared to control cells; moreover, titanium is osteoinductive per se on ASCs (+91 and +284% for ScI- and BFP-ASCs).
Finally, porcine autologous or heterologous sera, did not improve cell growth compared to standard condition.
No significant difference between cells from the different harvesting sites was observed, concerning all the features described.
In conclusions, swine buccal fat pad contains mesenchymal stem cells, and we suggest them for preclinical studies of periodontal and bone defects regeneration
17 β-estradiol (E2) differently affects osteogenic differentiation of mesenchymal stem cells from bone marrow and adipose tissue.
No full textEstrogens confer multiple skeletal protective effects both in vitro and in vivo, facilitating osteogenesis and suppressing osteoclast-induced bone resorption. Estrogen deficiency induces a modulation in osteoblasts and osteoclasts activity resulting in bone resorption increase. Besides, it also act on osteoblast precursors and bone marrow mesenchymal stem cells (BMSCs) impairing their proliferation and osteogenic activity. Mesenchymal stem cells (MSCs) are a heterogeneous population present in several tissues, and, due to their multipotency, low immunogenicity and no tumorigenicity they are an attractive tool in regenerative medicine. Although MSCs isolated from different tissues share common stemness features, recent findings show that they slightly differ in terms of gene and protein expression, suggesting an intrinsic difference among them. Since both BMSCs and ASCs (MSCs isolated from adipose tissue) are the most used in tissue engineering applied to bone regeneration, we evaluated in vitro the effect of 17β estradiol (E2) on their osteogenic differentiation ability. BMSCs and ASCs used in this study derived from two (♂, 55±16 y/o) and six (♂ and♀, 39±17y/o) patients, respectively. All the MSCs proliferated at constant rate when cultured till passage 10 with no changes in their morphology. In addition, at early passages, high clonogenicity (range 2.4-14.6%) and the typical mesenchymal stem cells immunophenotype were also determined. Additionally, osteogenic and adipogenic differentiation markers, such as ALP activity, collagen deposition and lipid vacuoles formation, became evident when properly induced. At first, MSCs treated with E2 (1, 10, 100, 1000 nM) for 14 days were as viable as control cells. Furthermore, 35 days treatment with E2 (10, 100nM) did not induce any modification in morphology and proliferation rate. In order to identify a pro-osteogenic effect of estradiol on MSCs, cells were pre-treated with 10, 100 nM E2 for 7, 21, 35 days and then both pre-treated and control cells were osteo-differentiated for 7 and 14 days either in the presence or in the absence of the hormone. Estradiol did not reduce their osteogenic differentiation; indeed cells differentiated with or without E2 significantly increased ALP activity and collagen deposition in comparison to control cells. Estradiol increased ALP activity in BMSCs by acting in synergy with osteogenic factors. The effect, already detectable in 7 days, was even more pronounced after 14 days of differentiation, with a significant increase of ALP activity of 49% and 67% in BMSCs incubated with E2 10 and 100nM, respectively, compared to untreated and differentiated cells. Interestingly, a significant dose-related increase of ALP activity was observed in differentiated (for 14 days) BMSCs pre-treated with the hormone for either 7 and 35 days, irrespective of the time of exposure. Surprisingly, E2 treatment never influenced the osteogenic differentiation ability of ASC at the doses and time points we have observed for BMSCs. In conclusion, E2 favoured osteogenic differentiation of just BMSCs and no effect on ASCs was observed, suggesting that either the hormone could activate different pathways or estrogen receptors are differently expressed in the two mesenchymal stem cell types
In vitro study on porcine Mesenchymal Stem Cells from Buccal Fat Pad as a novel population for periodontal regeneration: comparison with ASCs from subcutaneous tissue
No full textIntroduction
In our previous study we have characterized human mesenchymal stem cells from Bichat’s fat pad, and proposed them for future therapies of periodontal defects and oral bone regeneration [1]. Considering the swine an accepted model for preclinical study in tissue engineering applications, here we compare swine Adipose-derived Stem Cells (ASCs) from Buccal fat pad and from subcutaneous adipose tissue both alone and in combination with synthetic scaffolds
Materials and Methods
Cells isolated from subcutaneous interscapular site (ScI) and buccal fat pad (BFP) of 6 swine were characterized as previously described [1,2,3]. pASC interactions with titanium disk (Permedica S.p.A., Merate, Italy) and SIC–plasma-enhanced chemical vapour-deposition (SIC) fragments (CETEV – C.T.V. Carsoli - AQ, Italy) were also investigated, as well as cell growth in medium with porcine serum.
Results
5.5x104±3.3x104 and 3.0x104±9.3x103 cells/ml were isolated from interscapular subcutaneous adipose tissue (ScI) and buccal fat pad (BFP) (n=6), respectively. All the ASCs, with a fibroblast-like shape, costantly proliferated (DT of 82.9±11.5 and 72.5±8.2 hours for ScI- and BFP-ASCs, respectively) and showed a high clonogenic ability (+10.1±1.4% for ScI- and +8.9±1.5% for BFP-ASCs). FACS analysis revealed that both cell populations are similar in size and granularity, and are CD90+, whereas CD14, CD45 and CD271 were not detectable. Furthermore, after osteogenic induction, both ASCs increased either collagen and calcified extracellular matrix (ECM) production of about 87% and 118% for ScI-, and of about 254% and 116% for BFP-ASCs, respect to CTRL cells. Alkaline phosphatase activity (+126% and +201% for ScI- and BFP-ASCs, respectively) and osteonectin expression (+336% for ScI- and +306% for BFP-ASCs) were also up-regulated. In addition, both cell types were able to differentiate towards the adipogenic and chondrogenic lineages, as revealed by lipid vacuoles formation and GAGs deposition. Interestingly, the osteoinduction of ASCs in the presence of titanium and SIC-PECVD, increased calcified ECM depot compared to CTRL; in vitro titanium is also osteoinductive per se (+91 and +284% for ScI- and BFP-ASCs).
Finally, porcine sera, did not improve cell growth compared to the standard condition.
Discussion and Conclusions
No significant difference between stemness features and scaffold interactions of cells from the different harvesting sites was observed, suggesting that, as the human buccal fat pad (1), also porcine one contains mesenchymal stem cells. In conclusion, we propose porcine and human BFP-ASCs in allo- and xenogenic use in porcine model of periodontal diseases. Indeed, since the BFP is easily available for dentists and maxillofacial surgeons, these cells future use in these fields could be preferential.
References
1. Broccaioli et al, Biores Open Access 2(2):107-17 2013_2. de Girolamo et al, Cytotherapy 11(4):793-803 2009_3. Quirici et al, Stem Cell Dev 19(6):915-25 2010
Synthetic Bone Substitutes Differently Influence Human and Porcine Adipose-derived MSCs
No full textSwine are a proper animal model in bone tissue engineering for their similarities with humans in terms of bone healing, anatomy, and composition. Since cell-scaffold constructs are fundamental for bone regeneration, we studied the effects of two bone substitutes, RO-1 and RO-2, on human (h) and porcine (p) adipose-derived mesenchymal stem/stromal cells (ASCs), in vitro.
At first, we show that both cell types display stable proliferation rates, high clonogenicity, and an increase in ALP activity, collagen release, ECM calcification and osteopontin expression, when osteo-induced for 2 weeks.
The scaffolds are biocompatible, and both ASCs nicely adhere to them; however, while hASCs normally proliferate in the presence of biomaterials, a slight reduction in growth rate is observed for pASCs. Besides, ASC osteodifferentiation is influenced by biomaterials: RO-1 increased ALP activity and collagen release on human ASCs compared to cells in monolayer and seeded on RO-2, independently of the presence of osteogenic stimuli. Differently, pASCs cultured on scaffolds show 10 folds decrease of ALP activity compared to cells cultured in monolayer. In addition, RO-2 stimulates just ALP activity, while the presence of RO-1 induces mainly collagen production.
In conclusion, both biomaterials are biocompatible, however human and porcine ASCs interact differently with them suggesting the requirement of cell-specific in vitro tests before moving to in vivo model
Study of porcine adipose-derived stem cells for orthopaedic preclinical models
No full textIn this study we compare adipose-derived stem cells (ASCs) isolated from pigs and minipigs with human ASC (hASC) considering their use as a promising approach for osteochondral defects treatment to be tested in autologous preclinical model. ASCs, isolated from the interscapular region of both pigs (pASCs) and minipigs (mpASCs) and from patients undergone aesthetic liposuction, have been analysed for their clonogenicity, proliferation, osteogenic and chondrogenic potential. pASCs and mpASCs proliferate faster than human cells with a doubling time of about 66h for pASCs, 57h for mpASCs and 126h for hASCs. Porcine ASCs of both sources are more highly clonogenic with approximately 14% of clogenic cells until the passage 4 compared to human cells (3%). When porcine and human ASCs are induced to osteo-differentiate for 14 days, alkaline phosphatase (ALP) activity is upregulated of about 300% respect to undifferentiated cells; furthermore, an increase in collagene production of approximately 100% is also detected. Moreover, both chondrogenic differentiated pASCs and hASCs, aggregated into micromasses, express an abundant amount of Glycosaminoglycans (GAGs) showing a marked increase in comparison to undifferentiated cells; however, pASCs seem to require a modified differentiative condition since they suffer more than human cells when maintained in pellet culture. To better predict the outcome deriving from the use of porcine ASCs in bone defect models, we assess the osteogenic potential of these cells seeded on different biocompatible biomaterials including nanomodified titanium and collagen-hydroxyapatite composite scaffolds. In particular we observe an increase in ALP levels and collagene production both by undifferentiated and differentiated pASCs, mpASCs and hASCs. A synergistic effect produced by the presence of the scaffolds and the osteogenic stimuli is observed, supporting the possible use of ASCs bioconstructs in some future clinical applications. We conclude that pASCs, mpASCs and hASCs share in vitro some common features and possess a similar osteo-differentiative ability, supporting the idea that the pre-clinical autologous ASCs reimplantation model in pig or minipigs might be predictive of the behaviour of ASCs in future clinical applications of regenerative medicine
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