Non-critical Size Defects Research Articles

The effects of boron nitride/hydroxyapatite compounds on bone defects in osteoporotic rats

This study investigated the effects of poly-(lactide-co-glycolide) (PLGA) scaffolds containing different concentrations of boron nitride (BN) and/or hydroxyapatite (HA) on bone defects in osteoporotic rats. The control group consisted of healthy rats. A standard non-critical size defect was induced in the osteoporotic rat femurs 12 weeks post-ovariectomy. PLGA scaffolds containing different concentrations of BN+HA were then applied to the defect area. In one group, defect was induced but no PLGA was applied. Computed tomography images were obtained and tissue samples were collected in the first, second, and fourth weeks postoperatively. PLGA scaffolds were classified as no BN + HA, only 10% HA, 2.5% BN + 10% HA, 5% BN + 10% HA, 10% BN + 10% HA or only 2.5%, 5% or 10% BN. No healing was determined in the first and second weeks postoperatively. However, in the fourth week healing was observed in the groups treated with PLGA scaffolds containing 10% HA, 2.5% BN + 10% HA, 2.5% BN and 5% BN, and especially in the 2.5% BN + 10% HA group. The findings of this study suggest that BN may represent a novel target for treating osteoporotic bone defects for physicians and engineers.

Bioinformatic analysis of the molecular mechanisms underlying the progression of bone defects.

The pathophysiology of bone defects (BDs) is complex, and the treatment for bone defects, in particular massive bone defects, remains a major clinical challenge. Our study was conducted to explore the molecular events related to the progression of bone defects a common clinical condition. First, microarray data of GSE20980 were obtained from the Gene Expression Omnibus (GEO) database, where 33 samples in total were used to analyze the molecular biological processes related to bone defects. Next, the original data were normalized and differentially expressed genes (DEGs) were identified. Additionally, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted. Finally, a protein-protein interaction (PPI) network was constructed and the trends of the different genes were confirmed. Compared with the samples of non-critical size defects (NCSD), the samples of critical size defects (CSD) had 2057, 827, and 1,024 DEGs at 7, 14, and 21 days post injury, respectively. At day 7, the DEGs were significantly enriched in metabolic pathways, at day 14 the DEGs were predominantly enriched in G-protein coupled signaling pathways and the Janus kinase (JAK)-signal transducer and activator of transcription (STAT) signaling pathway, and at day 21 the DEGs were mainly enriched in circadian entrainment and synaptic-related functions. The PPI network showed similar results. Quantitative real-time PCR (qRT-PCR) and western blot (WB) were performed to validate the partial results of sequencing. This study provides some clues about the molecular mechanism behind bone defects, which should contribute to scientific research and clinical treatment of this condition.

Histological comparison of DBBM and platelet rich fibrin for guided bone regeneration in a rabbit model

To histologically evaluate the use of bovine derived deproteinized xenograft (DBBM), leukocyte and platelet rich fibrin (L-PRF) and the combination of both in Guided Bone Regeneration (GBR) performed in non-critical size defects in rabbit. A prospective experimental study was performed. Four bone defects in the tibiae of 12 rabbits were made and each of them was filled with DBBM, L-PRF, a combination of DBBM+L-PRF or was left to heal as control site. All defects were covered with a collagen membrane. Rabbits were randomly distributed in three groups and euthanatized at 3, 6 or 9 weeks. Samples were obtained and histologically analyzed to determine vital bone, connective tissue and remaining graft particles percentage. Analysis of variance, Kruskal Wallis and non-paired t-test where used to evaluate the significance of the results. At 3 weeks of healing, DBBM showed significantly more vital bone percentage than L-PRF (p=0,05) and DBBM+L-PRF showed significantly less connective tissue than control (p<0,05). All other groups showed no statistical difference between them. At 6 weeks of healing, DBBM showed significantly more vital bone percentage than L-PRF (p<0,05), DBBM+L-PRF (p<0,05) and control (p<0,05) and there wasn't any other significant difference regarding to connective tissue or remaining particle percentage between groups. At t 9 weeks healing period, there weren't any significant differences between groups. DBBM seems to enhance vital bone formation at early healing stages. The use of L-PRF alone or combined with DBBM, didn't show any histological improvement regarding to vital bone formation. The use of DBBM, alone or in conjunction with L-PRF showed a trend to reduce connective tissue percentage. The use of L-PRF combined with DBBM didn't affect the remaining particle percentage.

Scaffolds and coatings for bone regeneration.

Bone tissue has an astonishing self-healing capacity yet only for non-critical size defects (<6 mm) and clinical intervention is needed for critical-size defects and beyond that along with non-union bone fractures and bone defects larger than critical size represent a major healthcare problem. Autografts are, still, being used as preferred to treat large bone defects. Mostly, due to the presence of living differentiated and progenitor cells, its osteogenic, osteoinductive and osteoconductive properties that allow osteogenesis, vascularization, and provide structural support. Bone tissue engineering strategies have been proposed to overcome the limited supply of grafts. Complete and successful bone regeneration can be influenced by several factors namely: the age of the patient, health, gender and is expected that the ideal scaffold for bone regeneration combines factors such as bioactivity and osteoinductivity. The commercially available products have as their main function the replacement of bone. Moreover, scaffolds still present limitations including poor osteointegration and limited vascularization. The introduction of pores in scaffolds are being used to promote the osteointegration as it allows cell and vessel infiltration. Moreover, combinations with growth factors or coatings have been explored as they can improve the osteoconductive and osteoinductive properties of the scaffold. This review focuses on the bone defects treatments and on the research of scaffolds for bone regeneration. Moreover, it summarizes the latest progress in the development of coatings used in bone tissue engineering. Despite the interesting advances which include the development of hybrid scaffolds, there are still important challenges that need to be addressed in order to fasten translation of scaffolds into the clinical scenario. Finally, we must reflect on the main challenges for bone tissue regeneration. There is a need to achieve a proper mechanical properties to bear the load of movements; have a scaffolds with a structure that fit the bone anatomy.

Prx1 Expressing Cells Are Required for Periodontal Regeneration of the Mouse Incisor.

Previous studies have shown that post-natal skeletal stem cells expressing Paired-related homeobox 1 (PRX1 or PRRX1) are present in the periosteum of long bones where they contribute to post-natal bone development and regeneration. Our group also identified post-natal PRX1 expressing cells (pnPRX1+ cells) in mouse calvarial synarthroses (sutures) and showed that these cells are required for calvarial bone regeneration. Since calvarial synarthroses are similar to dentoalveolar gomphosis (periodontium) and since there is no information available on the presence or function of pnPRX1+ cells in the periodontium, the present study aimed at identifying and characterizing pnPRX1+ cells within the mouse periodontium and assess their contribution to periodontal development and regeneration. Here we demonstrated that pnPRX1+ cells are present within the periodontal ligament (PDL) of the mouse molars and of the continuously regenerating mouse incisor. By means of diphtheria toxin (DTA)-mediated conditional ablation of pnPRX1+ cells, we show that pnPRX1+ cells contribute to post-natal periodontal development of the molars and the incisor, as ablation of pnPRX1+ cells in 3-days old mice resulted in a significant enlargement of the PDL space after 18 days. The contribution of pnPRX1+ cells to periodontal regeneration was assessed by developing a novel non-critical size periodontal defect model. Outcomes showed that DTA-mediated post-natal ablation of pnPRX1+ cells results in lack of regeneration in periodontal non-critical size defects in the regeneration competent mouse incisors. Importantly, gene expression analysis of these cells shows a profile typical of quiescent cells, while gene expression analysis of human samples of periodontal stem cells (PDLSC) confirmed that Prx1 is highly expressed in human periodontium. In conclusion, pnPRX1+ cells are present within the continuously regenerating PDL of the mouse incisor, and at such location they contribute to post-natal periodontal development and regeneration. Since this study further reports the presence of PRX1 expressing cells within human periodontal ligament, we suggest that studying the mouse periodontal pnPRX1+ cells may provide significant information for the development of novel and more effective periodontal regenerative therapies in humans.

Absence of p21(WAF1/CIP1/SDI1) protects against osteopenia and minimizes bone loss after ovariectomy in a mouse model.

p21(WAF1/CIP1/SDI1) is a critical sentinel of the cell cycle that plays an important role in determining cell fate with respect to proliferation, differentiation and apoptosis. Recent studies have demonstrated that inhibition/loss of p21 promotes osteo-chondro differentiation in progenitor/stem cells, and that p21 knockout (p21-/-) mice demonstrate enhanced bone regeneration compared to wild-type controls after a non-critical size defect. It was therefore hypothesized that the absence of p21 may also protect against bone loss through enhancing bone formation, tilting the balance away from bone resorption, in an ovariectomy-induced osteopenia mouse model, investigated via microCT imaging. While p21-/- mice demonstrated significantly less bone loss after ovariectomy compared to wild-type controls, no increase in the number osteoclasts or osteoblasts in the bone or bone marrow was observed, nor was there an increase in osteoclast activity. Therefore, while the absence of p21 protected mice against estrogen mediated bone loss, the mechanisms/pathways responsible remained elusive. This study demonstrates that p21 may play a significant role in bone remodeling, and a better understanding of how the p21 pathway regulates bone anabolism and catabolism could lead to novel therapies for osteoporosis in the future.

The Obliteration of Noncritical Size Bone Defects With Bone Dust or Bone Replacement Material (Bioactive Glass S53P4).

Bone dust (BD) harvested during operation may be suitable as an autologous obliteration material for noncritical size defects. Bioactive glass (BA) can be an alternative. To treat noncritical size defects, BD and BA are commonly used for obliteration techniques. However, the optimal harvesting method and parameters for BD have not been examined. In this study, we analyzed the osseoregenerative potential of both materials. Thirteen female merino sheep (7-yr old) underwent surgery on the frontal calvaria. Three defects were inserted. The first defect was considered a reference and remained unfilled, the second defect was filled with BD from the calvaria bone, and the third defect was filled with BA S53P4. The animals were sacrificed after 3 weeks. To evaluate bone regeneration, we used digital volume tomography, bone density measurement, fluorochrome sequence labeling, and histological analysis. All analyses showed quantitative and qualitative bone regeneration 3 weeks after operation. The control blank defect showed significantly less new bone growth than the BD-filled defect. Moreover, bone regeneration occurred from the surrounding bone and showed only a defect bridge in the BD-filled defect. The BA completely filled the defect and had the highest density although the same amount of new mineralized bone generated as in the reference. BD and BA seemed to be suitable bone replacement materials for obliteration techniques because they completely filled the defects. Thus, BD harvested under standardized conditions provided a higher level of osteoreparation potential for the generation of woven bone and establishment of defect bridges.

Lattice Microarchitecture for Bone Tissue Engineering from Calcium Phosphate Compared to Titanium.

Additive manufacturing of bone tissue engineering scaffolds will become a key element for personalized bone tissue engineering in the near future. Several additive manufacturing processes are based on extrusion where the deposition of the filament will result in a three-dimensional lattice structure. Recently, we studied diverse lattice structures for bone tissue engineering realized by laser sintering of titanium. In this work, we used lithography-based ceramic manufacturing of lattice structures to produce scaffolds from tricalcium phosphates (TCP) and compared them in vivo to congruent titanium scaffolds manufactured with the identical computer-aided design data to look for material-based differences in bony healing. The results show that, during a 4-week period in a noncritical-size defect in a rabbit calvarium, both scaffolds with the identical microarchitecture performed equally well in terms of bony regeneration and bony bridging of the defect. A significant increase in both parameters could only be achieved when the TCP-based scaffolds were doped with bone morphogenetic protein-2. In a critical-size defect in the calvarial bone of rabbits, however, the titanium scaffold performed significantly better than the TCP-based scaffold, most likely due to its higher mechanical stability. We conclude that titanium and TCP-based scaffolds of the same microarchitecture perform equally well in terms of bone regeneration, provided the microarchitecture meets the mechanical demand at the site of implantation.

Impairment of the chondrogenic phase of endochondral ossification in vivo by inhibition of cyclooxygenase-2.

Many studies have reported on the effects of cyclooxygenase-2 (COX-2) inhibition on osteogenesis. However, far less is known about the effects of COX-2 inhibition on chondrogenic differentiation. Previous studies conducted by our group show that COX-2 inhibition influences in vitro chondrogenic differentiation. Importantly, this might have consequences on endochondral ossification processes occurring in vivo, such as bone fracture healing, growth plate development and ectopic generation of cartilage. The goal of our study was to investigate, in vivo, the effect of COX-2 inhibition by celecoxib on the cartilaginous phase of three different endochondral ossification scenarios. 10 mg/kg/day celecoxib or placebo were orally administered for 25 d to skeletally-immature New Zealand White rabbits (n = 6 per group). Endochondral ossification during fracture healing of a non-critical size defect in the ulna, femoral growth plate and ectopically-induced cartilaginous tissue were examined by radiography, micro-computed tomography (µ-CT), histology and gene expression analysis. Celecoxib treatment resulted in delayed bone fracture healing, alterations in growth plate development and progression of mineralisation. In addition, chondrogenic differentiation of ectopically-induced cartilaginous tissue was severely impaired by celecoxib. In conclusion, we found that celecoxib impaired the chondrogenic phase of endochondral ossification.

The effect of L-PRF membranes on bone healing in rabbit tibiae bone defects: micro-CT and biomarker results

More insight into the biological fundamentals of leukocyte platelet-rich fibrin (L-PRF) guided healing is necessary to recommend its application, in particular in deficient bone sites that need to support implants. This study investigated the short-term bone healing effect of L-PRF treatment in cylindrical non-critical sized bone defects with 3 mm diameter and 6 mm depth in tibiae of 18 adult male New Zealand White rabbits. After a randomization process, 96 bone defects were prepared and half of them were filled with a L-PRF membrane, while untreated defects in the opposite tibia served as control group. The rabbits were euthanized after 7, 14 or 28 days of healing. The bone healing of the cortical and medullary areas was investigated by micro-CT, while the expression of molecular markers (RUNX2, VEGFA, COL1A2 and BMP2) was assessed by qRT-PCR. Treatment with L-PRF did not affect the micro-structural bone characteristics of the repaired bone tissue, except for a decrease in the trabecular connectivity at the cortical level after 14 days of healing. At this time, RUNX2 and VEGFA mRNA levels were significantly lower in the treated defects. L-PRF membranes thus had a temporary negative influence on the bone microarchitecture (Tb.Pf) and on the RUNX2 and VEGFA expression during early bone healing. Overall, L-PRF treatment did not enhance bone regeneration in these non-critical size defects after 28 days.

Osteoconduction in keratin-hydroxyapatite composite bone-graft substitutes.

Reconstituted keratin-hydroxyapatite (K-HA) composites have shown potential as nonload-bearing bone graft substitute material. This in vivo study investigated the bone regeneration response of keratin plus 40% HA composite materials in comparison to collagen counterparts and an unfilled defect site. The implantation site was a noncritical size defect created in the long bones (tibia) of sheep, with observations made at 1, 2, 4, 6, 8, and 12weeks postimplantation. Porous K-HA materials displayed an excellent biocompatibility similar to collagen counterparts; however, the rate of bone regeneration at K-HA implantation sites was markedly slower than that of the collagen or unfilled defect sites. While collagen materials were undetectable by 4weeks implantation, K-HA composite remnants were present at 12weeks. However, there is evidence that K-HA implants participated in the natural remodelling process of bone, with bone regeneration occurring via a creeping substitution mechanism. Observations imply that the rate of bone ingrowth into the K-HA defect site was matched with the rate of K-HA resorption. These results suggest that K-HA materials may offer significant benefits as nonload-bearing bone graft substitutes where it is desirable that the degradation of the scaffolding material be well matched with the rate of bone regeneration. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 2034-2044, 2017.

Evaluation of tensile strength resistance of different biomaterials in calvarial rats

AIM: The aim of this study was to determine the effect of different grafting materials on bone tensile strength after 6-month wound healing. MATERIAL AND METHODS: Non-critical size defects (3-mm diameter) were created in calvarium of 30 three-month-old Wistar rats. Animals were divided into 3 groups (n=10) treated with different grafting materials: GenOx® (Group 1A); GenMix® (Group 1B); no treatment (blood clot, Group 3). Six months after the surgery, rats were sacrificed; bone specimens were harvested and submitted to tensile strength test using a universal testing machine. The bone fracture surface morphology was evaluated using scanning electron microscopy (SEM) at 200X magnification. Data were compared by One-Way ANOVA at 5% significance. RESULTS: No significant difference was found among the groups although tensile strength decreased in the following order: Group 3 (9.56±3.74MPa), Group 1B (8.58±3.60MPa), Group 1A (7.70±2.41MPa). All tested materials showed similar effects on bone tensile strength, no matter the source (xenogenic or blood clot). CONCLUSION: After six months of bone healing, the type of grafting material is irrelevant to the final outcome and bone tensile strength.

Comparative evaluation of three calcium phosphate synthetic block bone graft materials for bone regeneration in rabbit calvaria

Various synthetic materials were developed to be used for implant dentistry and periodontal treatments. Among the various synthetic bone substitutes, calcium phosphate ceramics have been extensively investigated because their mineral chemistry resembles that of human bone. We evaluated the regeneration of three calcium phosphate synthetic block bone grafts in rabbit calvarial noncritical size defects. Four 8-mm-diameter defects were created in each rabbit (N = 10). Three defects, hydroxyapatite (HA), beta-tricalcium phosphate (β-TCP), and biphasic calcium phosphate (BCP), were randomly filled with one of three fabricated synthetic block bone graft materials. The fourth defect was filled with blood clots. Specimens were harvested at 4 and 8 weeks postsurgery. Histological and histometrical findings indicated that all three calcium phosphate block bone graft materials were able to maintain space significantly better than the control group at both 4 and 8 weeks. In the BCP group, the amount of newly formed bone was increased more than for the other groups. Additionally, β-TCP showed a large resorption of graft materials after 4 weeks postsurgery, while there were only small resorption for HA and BCP.

Removal of fracture hematoma and replacement with fresh hematoma delays bone healing in sheep

Removal of fracture hematoma and replacement with fresh hematoma delays bone healing in sheep

Bone formation in TiO2 bone scaffolds in extraction sockets of minipigs

The osteoconductive capacity of TiO2 scaffolds was investigated by analysing the bone ingrowth into the scaffold structure following their placement into surgically modified extraction sockets in Gottingen minipigs. Non-critical size defects were used in order to ensure sufficient bone regeneration for the evaluation of bone ingrowth to the porous scaffold structure, and sham sites were used as positive control. Microcomputed tomographic analysis revealed 73.6±11.1% of the available scaffold pore space to be occupied by newly formed bone tissue, and the volumetric bone mineral density of the regenerated bone was comparable to that of the native cortical bone. Furthermore, histological evidence of vascularization and the presence of bone lamellae surrounding some of the blood vessels were also observed within the inner regions of the scaffold, indicating that the highly interconnected pore structure of the TiO2 scaffolds supports unobstructed formation of viable bone tissue within the entire scaffold structure. In addition, bone tissue was found to be in direct contact with 50.0±21.5% of the TiO2 struts, demonstrating the good biocompatibility and osteoconductivity of the scaffold material.

Dimensional Ridge Preservation with a Novel Highly Porous TiO2Scaffold: An Experimental Study in Minipigs

Despite being considered noncritical size defects, extraction sockets often require the use of bone grafts or bone graft substitutes in order to facilitate a stable implant site with an aesthetically pleasing mucosal architecture and prosthetic reconstruction. In the present study, the effect of novel TiO2 scaffolds on dimensional ridge preservation was evaluated following their placement into surgically modified extraction sockets in the premolar region of minipig mandibles. After six weeks of healing, the scaffolds were wellintegrated in the alveolar bone, and the convex shape of the alveolar crest was preserved. The scaffolds were found to partially preserve the dimensions of the native buccal and lingual bone walls adjacent to the defect site. A tendency towards more pronounced vertical ridge resorption, particularly in the buccal bone wall of the nongrafted alveoli, indicates that the TiO2 scaffold may be used for suppressing the loss of bone that normally follows tooth extraction.

Effect of Carbonate-Apatite on Bone Repair in Non-Critical Size Defect of Rat Calvaria

The objective of this study was to investigate the bone repair of carbonate apatite (cHA) in comparison to hydroxyapatite (HA, control group), on osseous repair of non-critical size defect in rat calvaria. Spheres (400&lt;ø&gt;500μm) of both materials were synthesized under 37°C (cHA) and 90°C (HA) and were not sintered. Fifteen rats Wistar were submitted to general anesthesia and two perforations (4mm each) were made, one in each parietal bone, for implantation of cHA (left side) and HA spheres (right side). After 1, 3 and 6 weeks, five animals of each group were killed and the two fragments with the biomaterial were collected from the calvaria. The bone blocks with biomaterial were demineralized and 5μm thick semi-serial sections were done for histological analysis. The experimental group of 6 weeks did not show the presence of spheres of both biomaterials and few spheres were observed after 1 and 3 weeks. Histological analysis showed the connective tissue repairing the surgical defect after 1 week and newly formed bone after 3 weeks of surgery. Thus, we concluded both materials are biocompatible, promote osteoconduction and in all studied periods the biomaterials showed to be resorbable.

Human adipose derived stem cells reduce callus volume upon BMP-2 administration in bone regeneration

IntroductionThe demand for new therapeutic approaches to treat bone defects and fractures is increasing in trauma surgery and orthopaedics because the number of patients with degenerative diseases is continuously growing. “Tissue Engineering” offers promising new technologies that combine the three components – cells, growth factors and matrix. Efforts are targeted at improving and accelerating recovery, especially for long bone fractures, and reducing the risk of delayed bone healing or pseudoarthrosis. Adult human adipose-derived stem cells (ASC) can differentiate into osteoblasts in an osteogenic surrounding. Bone morphogenetic protein-2 (BMP-2) accelerates and initiates this differentiation. Fibrin, a matrix that promotes wound healing, is a promising carrier for ASCs and BMP-2. Materials and methodsIn this study, a 2mm transcortical drill hole in the femur of male rats served as a small non-critical size defect model for fracture simulation. In vivo bone healing was investigated upon administration of the growth factor BMP-2 embedded with ASCs in a locally applied fibrin matrix. Groups with the components alone were also investigated. After 2 and 4 weeks, μCT and histology were performed to determine the bone and callus volume. Results and discussionAfter only a short period of time (2 and 4 weeks), this animal model discloses comparative information about the osteogenetic potential and bone regeneration with little effort (no osteosynthesis necessary). The most significant result found in this model is that the combination of ASCs and BMP-2 in a fibrin matrix significantly reduces callus formation after 2 weeks compared to BMP-2 alone. BMP-2 alone significantly increased callus formation. ASCs embedded alone in the fibrin matrix did not lead to increased bone regeneration. ConclusionTransplantation of ASC modulated the callus induction by BMP-2 to a normal volume.

Bone regeneration in the presence of a synthetic hydroxyapatite/silica oxide-based and a xenogenic hydroxyapatite-based bone substitute material

A comparison of synthetic hydroxyapatite/silica oxide, xenogenic hydroxyapatite-based bone substitute materials with empty control sites in terms of bone regeneration enhancement in a rabbit calvarial four non-critical-sized defect model. In each of six rabbits, four bicortical calvarial bone defects were generated. The following four treatment modalities were randomly allocated: (1) empty control site, (2) synthetic hydroxyapatite/silica oxide-based (HA/SiO) test granules, (3) xenogenic hydroxyapatite -based granules, (4) synthetic hydroxyapatite/silica oxide -based (HA/SiO) test two granules. The results of the latter granules have not been reported due to their size being three times bigger than the other two granule types. After 4 weeks, the animals were sacrificed and un-decalcified sections were obtained for histological analyses. For statistical analysis, the Kruskal-Wallis test was applied (P<0.05). Histomorphometric analysis showed an average area fraction of newly formed bone of 12.32±10.36% for the empty control, 17.47±6.42% for the xenogenic hydroxyapatite -based granules group, and 21.2±5.32% for the group treated with synthetic hydroxyapatite/silica oxide -based granules. Based on the middle section, newly formed bone bridged the defect to 38.33±37.55% in the empty control group, 54.33±22.12% in the xenogenic hydroxyapatite -based granules group, and to 79±13.31% in the synthetic hydroxyapatite/silica oxide -based granules group. The bone-to-bone substitute contact was 46.38±18.98% for the xenogenic and 59.86±14.92% for the synthetic hydroxyapatite/silica oxide-based granules group. No significant difference in terms of bone formation and defect bridging could be detected between the two bone substitute materials or the empty defect. There is evidence that the synthetic hydroxyapatite/silica oxide granules provide comparable results with a standard xenogenic bovine mineral in terms of bone formation and defect bridging in non-critical size defects.

The performance of poly- ε-caprolactone scaffolds in a rabbit femur model with and without autologous stromal cells and BMP4

The ability of a cellular construct to guide and promote tissue repair strongly relies on three components, namely, cell, scaffold and growth factors. We aimed to investigate the osteopromotive properties of cellular constructs composed of poly- ε-caprolactone (PCL) and rabbit bone marrow stromal cells (BMSCs), or BMSCs engineered to express bone morphogenetic protein 4 (BMP4). Highly porous biodegradable PCL scaffolds were obtained via phase inversion/salt leaching technique. BMSCs and transfected BMSCs were seeded within the scaffolds by using an alternate flow perfusion system and implanted into non-critical size defects in New Zealand rabbit femurs. In vivo biocompatibility, osteogenic and angiogenic effects induced by the presence of scaffolds were assessed by histology and histomorphometry of the femurs, retrieved 4 and 8 weeks after surgery. PCL without cells showed scarce bone formation at the scaffold–bone interface (29% bone/implant contact and 62% fibrous tissue/implant contact) and scarce PCL resorption (16%). Conversely, PCL seeded with autologous BMSCs stimulated new tissue formation into the macropores of the implant (20%) and neo-tissue vascularization. Finally, the BMP4-expressing BMSCs strongly favoured osteoinductivity of cellular constructs, as demonstrated by a more extensive bone/scaffold contact.