In vivo comparison of hard tissue regeneration with ovine mesenchymal stem cells processed with either the FICOLL method or the BMAC method.

INTRODUCTION: The aim of this work was to analyse the suitability of mesenchymal stem cell isolation by FICOLL density centrifugation and the closed bone marrow aspirate concentrate (BMAC) system for sinus augmentation with bovine bone mineral (BBM) in the sheep model. METHODS: 16 sheep underwent sinus augmentation with BBM and MSCs; they were divided between two groups with survival points of 8 and 16 weeks. For the FICOLL control arm three, and for the BMAC test arm, five augmentations were performed for each time point. The derived cell numbers were counted; a colony forming unit (CFU) assay was performed; the pluripotency of the MSCs was proved; histological and histomorphometrical analysis were performed. RESULTS: The approach of using BBM and MSCs in combination with fibrin adhesive was sufficient for new bone formation as the FICOLL experiment indicated. However, due to significantly lower cell numbers isolated using the BMAC in sheep, less new bone was formed in the test arm. CONCLUSIONS: The BMAC system is well suited for human MSC isolation but it needs to be optimized to fit sheep cell characteristics if it is to be used in this animal model.

Maxillary sinus floor elevation surgery with BioOss® mixed with a bone marrow concentrate or autogenous bone: test of principle on implant survival and clinical performance.

The purpose of this study was to assess implant survival and 1-year clinical performance of implants placed in the posterior maxilla that had been subjected to maxillary sinus floor elevation surgery with bovine bone mineral (BioOss®) mixed with autogenous bone marrow concentrate or autogenous bone. In a randomized, controlled, split-mouth design study, a bilateral sinus floor augmentation procedure was performed in 12 edentulous patients. At random, one side was treated with BioOss® seeded with an iliac crest bone marrow concentrate enriched in mesenchymal stem cells (test side) and the other with BioOss® mixed with autogenous bone (control side). Three to four months after augmentation, 66 implants were placed. Implant survival, plaque, gingival, and bleeding indices, probing depth, and peri-implant radiographic bone levels were assessed at baseline and 12 months after functional loading. During osseointegration, three implants failed on the test side (two patients) and no implants failed on the control side, resulting in 3-month survival rates of 91% and 100%, respectively. No implants were lost after functional loading and no differences in soft tissue parameters or peri-implant bone loss were observed between the control and test sides. After 1 year in function, no clinically relevant differences were observed regarding soft tissue parameters or peri-implant bone loss.

Environmental biomechanics substantiated by defined pillar micropatterns govern behavior of human mesenchymal stem cells.

While evidence on the impact of the biomechanical environment elasticity on human mesenchymal stem cell (hMSC) behavior is growing, the aspect of micropatterning is still poorly understood. Thus, the present study aimed at investigating the influence of defined environmental micropatterning on hMSC behavior. Following characterization, hMSCs were grown on defined pillar micropatterns of 5, 7, 9, and 11 µm. With respect to cell behavior, primary hMSC adhesion was detected by indirect immunofluorescence (iIF) for paxillin, vinculin, integrin αV, and actin, while proliferation was visualized by histone H3. Morphogenesis was monitored by scanning electron microscopy and the expression of stem cell-specific biomarkers by real-time PCR. Favoritism of primary adhesion of hMSCs on pillar tops occurred at smaller pillar micropatterns, concomitant with cell flattening. While vinculin, integrin αV, and paxillin appeared initially more cytoplasmic, high pillar micropatterns favored a progressive redistribution with polarization to cell tension sites and at cell borders. Accomplishment of morphogenesis at day 3 revealed establishment of fully rotund cell somata at 5 µm, while hMSCs appeared progressively elongated at rising micropatterns. The hMSC proliferation capacity was influenced by pillar micropatterns and gene expression analysis of stem cell- and differentiation-associated biomarkers disclosed clear modulation by distinct pillar micropatterns. In response to environmental biomechanics, our results show that hMSC behavior is governed by pillar micropatterning. In turn, these findings may form the basis to prospectively direct lineage specificity of hMSCs in a customized fashion.

Applicability of equine hydroxyapatite collagen (eHAC) bone blocks for lateral augmentation of the alveolar crest. A histological and histomorphometric analysis in rats.

This study assessed the mechanical characteristics, biocompatibility and osteoconductive properties of an equine hydroxyapatite collagen (eHAC) bone block when applied as a bone substitute for lateral augmentation of rat mandible. 96 rats underwent lateral augmentation of the mandible, using two substitute bone blocks (eHAC or Bio-Oss spongiosa) or autologous bone grafts. Signs of inflammation, amount of bone formation and ingrowth of bone into the bone blocks were assessed at 1 and 3 months. eHAC blocks were mechanically rigid and could be fixed firmly and easily. Bio-Oss spongiosa blocks were brittle and fixation was difficult. eHAC and Bio-Oss spongiosa blocks were biocompatible and induced few or no signs of inflammation. Inflammation prevalence between the groups was not statistically different. Bone formation and bone growth into the blocks was significantly higher in eHAC than Bio-Oss spongiosa blocks, but lower than in autologous bone grafts (after 1 and 3 months). Regression analysis showed that the autologous bone graft predicted new bone formation at both time points. The eHAC block was only a predictor at 1 month; a trend was found at 3 months. The application of biodegradable membranes was not related to more bone ingrowth.

Osteopenia in the maxillofacial area: a study in sheep.

UNLABELLED: Osteoporosis is a major public health problem worldwide. Its significance in the fields of traumatology and implantology of the maxillofacial area requires investigation. A large animal model was used to assess bone loss in the lumbar spine and mandible. INTRODUCTION: Osteoporosis is a prevalent disease characterized by low bone mass and microarchitectural deterioration of bone tissue. Osteoporosis-related fractures represent a major public health burden. The presence and relevance of osteoporosis in the maxillofacial area remain controversial. Research in humans is limited by difficulties in finding large homogenous study groups and, due to ethical considerations, numerous animal models have been used in osteoporosis research. The aim of this study was to assess a sheep model of generalized osteopenia for changes in the maxillofacial area. METHODS: Bone loss was induced in ten Merino sheep by ovariectomy, intramuscular administration of glucocorticoids, and a calcium-reduced diet. Five untreated animals served as controls. Bone mineral density (BMD) was assessed by dual-energy X-ray absorptiometry at the lumbar spine at baseline and after 1, 3, 6, 9, and 12 months. Lumbar and mandibular bone biopsies were obtained and analyzed with microcomputed tomography. RESULTS: Lumbar BMD decreased progressively in the intervention group and was most significantly low after 6 months (p < 0.001). Lumbar trabecular bone showed a significant decrease in bone volume (BV)/tissue volume (TV; p < 0.05) in the inducted group. Significant changes were found in both analyzed mandibular regions for BV/TV (p < 0.05). Regional variations were found for other parameters in the mandible. The cortical width was substantially reduced in the intervention group (p < 0.001). CONCLUSIONS: Microstructural changes occurring in sheep as a result of induction seem to have a generalized nature. This sheep model meets the criteria for further investigation in the maxillofacial area.

Maxillary sinus floor elevation with bovine bone mineral combined with either autogenous bone or autogenous stem cells: a prospective randomized clinical trial.

AIM: To assess whether differences occur in bone formation after maxillary sinus floor elevation surgery with bovine bone mineral (BioOss(®)) mixed with autogenous bone or autogenous stem cells. The primary endpoint was the percentage of new bone three months after the elevation procedure. MATERIAL AND METHODS: In a randomized, controlled split-mouth design, in 12 consecutive patients (age 60.8 ± 5.9 years, range 48-69 years) needing reconstruction of their atrophic maxilla, a bilateral sinus floor augmentation procedure was performed. Randomly, on one side the augmentation procedure was performed with bovine bone mineral (BioOss(®)) seeded with mononuclear stem cells harvested from the posterior iliac crest (test group) while BioOss(®) mixed with autogenous bone (harvested from the retromolar area) was applied on the contra-lateral side (control group). On 14.8 ± 0.7 weeks after the sinus floor elevation, biopsies from the reconstructed areas were taken at the spots where subsequently the endosseous implants were placed. The biopsies were histomorphometrically analyzed. RESULTS: Significantly more bone formation was observed in the test group (17.7 ± 7.3%) when compared with the control group (12.0%± 6.6; P=0.026). In both the test and control group, all implants could be placed with primary stability. In one patient, not all biopsies contained BioOss(®). This patient was excluded from analysis. CONCLUSION: Mesenchymal stem cells seeded on BioOss(®) particles can induce the formation of a sufficient volume of new bone to enable the reliable placement of implants within a time frame comparable with that of applying either solely autogenous bone or a mixture of autogenous bone and BioOss(®). This technique could be an alternative to using autografts.

Miniplate osteosynthesis with four different systems in sheep.

The aim of this study was to compare a combination of a locking system with self-tapping (ST-L) or self-drilling-tapping (SDT-L) screws with a combination of conventional miniplates with self-tapping (ST) and self-forming (SF) screws. A standardized osteotomy and osteosynthesis with one of the above mentioned systems was performed in 24 sheep. Callus formation was measured with the help of CT scans assisted by a navigation system. Specimens of each osteotomy gap were taken and examined histologically. The best results were observed when self-tapping screws and the Mini-Locking-System (ST-L) were applied. The slowest healing was seen in animals treated with miniplates and SF screws. After 8 weeks an increase in bone formation could be observed in the ST, SF, SDT-L systems. The results after 8 weeks were comparable with those achieved by the ST-L system after 4 weeks. The improved stability of the osteosynthesis with the ST-L system resulted in early ossification of the osteotomy gap and the smallest amount of callus formation.