Comparison of rabbit rib defect regeneration with and without graft.

Rib segment, as one of the most widely used autologous boneresources for bone repair, is commonly isolated with an empty left in the defect. Although defective rib repair is thought to be unnecessary traditionally, it's of vital importance actually to promote rib regeneration for patients with better postoperative recovery and higher life quality. Comparative investigations on rabbit rib bone regeneration with and without graft were reported in this article. A segmental defect was performed on the 8th rib of 4-month-old male New Zealand rabbits. The mineralized collagen bone graft (MC) was implanted into the defect and evaluated for up to 12 weeks. The rib bone repair was investigated by using X-ray at 4, 8 and 12 weeks and histological examinations at 12 weeks after surgery, which showed a higher bone remodeling activity in the groups with MC implantation in comparison with blank control group, especially at the early stage of remodeling.

Rib Composite Flap With Intercostal Nerve and Internal Thoracic Vessels for Mandibular Reconstruction.

PURPOSE: The purpose of this study was to present the outcome and discuss the feasibility of rib composite flap with intercostal nerve and internal thoracic vessels for reconstructing mandibular defect. METHODS: Rib composite flaps have been used in 82 patients for reconstructing benign tumor-caused large mandibular defects: 66 of the 82 patients were reconstructed using rib composite flap with intercostal nerve and internal thoracic vessels, whereas the other 16 patients were reconstructed using rib composite flap with internal thoracic vessels, without intercostal nerve. After operation, clinical observation, imageological examination, and sensory detection were used to evaluate the effect of reconstruction. RESULTS: All rib composite flaps with intercostal nerve and internal thoracic vessels were successfully harvested and transplanted. Both immediate and long-term examination showed good appearance reconstruction. All followed-up patients conveyed good satisfaction degree with function and appearance reconstruction. Postoperative panoramic x-ray examination showed new bone formation between the transplanted rib and mandibular stump. Good recoveries of mandibular nerve sensory were observed when followed up after reconstruction surgery. CONCLUSIONS: Rib composite flap with intercostal nerve and internal thoracic vessels could be a promising method for reconstruction of mandibular defects.

Combination of hyaluronic acid hydrogel scaffold and PLGA microspheres for supporting survival of neural stem cells.

PURPOSE: To develop a biomaterial composite for promoting proliferation and migration of neural stem cells (NSCs), as well as angiogenesis on the materials, to rescue central nervous system (CNS) injuries. METHODS: A delivery system was constructed based on cross-linked hyaluronic acid (HA) hydrogels, containing embedded BDNF and VEGF-loaded poly(lactic-co-glycolic acid) (PLGA) microspheres for controlled delivery and support for NSCs in the CNS. The surface morphologies were evaluated by SEM and AFM, mechanical property was investigated by rheological tests, and release kinetics were performed by ELISA. Bioactivity of released BDNF and VEGF was assessed by neuron and endothelial cell culture, respectively. Compatibility with NSCs was studied by immunofluorescent staining. RESULTS: Release kinetics showed the delivery of BDNF and VEGF from PLGA microspheres and HA hydrogel composite were sustainable and stable, releasing ~20-30% within 150 h. The bioactivities preserved well to promote survival and growth of the cells. Evaluation of structure and mechanical properties showed the hydrogel composite possessed an elastic scaffold structure. Biocompatibility assay showed NSCs adhered and proliferated well on the hydrogel. CONCLUSIONS: Our created HA hydrogel/PLGA microsphere systems have a good potential for controlled delivery of varied biofactors and supporting NSCs for brain repair and implantation.

The mechanical anisotropy on a longitudinal section of human enamel studied by nanoindentation.

Nanoindentation has been widely used for probing the mechanical properties of tooth, especially for characterizing its complex hierarchical structures. Previous studies have confirmed the anisotropic mechanical behaviors caused by the alternated orientations of enamel rods and the alignment of fibril-like hydroxyapatite crystals, but the longitudinal section of enamel, which was composed of parallel-arranged rods, was regarded as a homogeneous continuum as always. In this study, nanoindentation combined with SEM was carried out with the indenter rotating on the longitudinal section of enamel to evaluate the relativity between the nano-mechanical properties and the orientation of indentation impressions. It has been shown that the enamel presented different elastic modulus and hardness with different angles of indenter on its longitudinal section, and its anisotropy was also confirmed by the remarkable asymmetric morphologies of impressions. We observed that the parallel arrangement of crystal fibrils and enamel rods might trigger the expansion of the micro-cracks in preferred orientation, and result in scalene triangle indentation impressions, altering contact areas as well as inconsistent mechanical behaviors. Consequently, it is considered that the longitudinal sections of enamel should be modeled as anisotropic.

[Biocompatible study of modified titanium skirt for keratoprosthesis].

OBJECTIVE: The research is engaged in developing an improved titanium skirt for keratoprosthesis, the aim of this study was to evaluate biocompatible of keratoprosthesis of novel design. METHODS: The pure titanium skirt for keratoprosthesis with three hands were first Sandblasted, and then bioactive hydroxyapatite coated on Sandblasted titanium for keratoprosthesis by a acid-alkali chemical treatment. A total of 18 New Zealand white rabbits and 18 alkali burned rabbit corneas were respectively divided into three groups (Group A, B and C; Group E, F and G) with simple random sampling methods. The modified titanium skirt was inserted into the corneal stroma of Group A and E, and then the control skirt was inserted into Group B and F. Group C and H did not insert skirt as surgery control. The interfacial biointegration of skirt/cornea were examined under light microscopy by HE, TEM and SEM. The transparent center was implanted to rabbit corneas with modified titanium skirt after 3 months. RESULTS: Dense hydroxyapatite coating was deposited on the Sandblasted specimens by an acid-alkali chemical treatment. The number of corneal fibroblasts increased significantly in Group A compared with Group B. The extracellular matrix deposited on the surface of modified titanium skirt was more dense and tight than that of control skirt. There was a significant difference in the shear force of skirt among groups A and B (t = 3.297, P < 0.05), E and F (t = 4.237, P < 0.05), and taking out the skirt in Group A and E were more difficulty than that in Group B and F after 3 months. The observation of the tissue sections of modified skirt inserted eyes revealed that there were cells and collagen-like fibres perpendicular to or at an angle to the rough interface and the cellular function was extremely active, in accordance with the results of TEM. Compared with healthy host tissue, skirt/cornea healing after alkali burn was impaired. CONCLUSIONS: Hydroxyapatite modified Sandblasted titanium skirt for keratoprosthesis can promote the interfacial biointegration of skirt and host cornea, no matter in healthy cornea or diseased cornea (alkali burn cornea). Hydroxyapatite coating improved the bioactivity of titanium.

Disturbed enamel biomineralization in col1-caPPR mouse incisor.

During the mineralization process of enamel, gene expression controls the activities of ameloblasts, the secretion and assembly of an extracellular protein matrix, affecting the final structure and functions. In this study, the enamel in the maxillary and mandibular incisors of wild-type and transgenic (col1-caPPR) mice, in which a constitutively active PTH/PTHrP receptor (PPR) was targeted to osteoblastic cells, was observed by scanning electron microscopy (SEM), Fourier transform infrared microscopy (FTIRM), and nanoindentation. The SEM studies showed that several different patterns of aberrations in crystal arrangement, disturbed prism organization without decussation, as well as abnormal enamel distribution were encountered in transgenic enamel. FTIRM analysis revealed poorer crystallinity/maturity after mutation. Nanoindentation measurement disclosed that transgenic enamel had 24.6% lower hardness and 12.3% lower elastic modulus. We attributed the inferior properties to the loosely packing crystals and abnormal prism organization. Furthermore, the col1-caPPR mouse model was substantiated to be useful to study how genes modulate the biomineralization process.

Therapeutic effects of hepatocyte growth factor-overexpressing dental pulp stem cells on liver cirrhosis in a rat model.

Cirrhosis is the terminal stage of hepatic diseases and is prone to develop into hepatocyte carcinoma. Increasing evidence suggests that the transplantation of dental pulp stem cells (DPSCs) may promote recovery from cirrhosis, but the key regulatory mechanisms involved remain to be determined. In this study, we overexpressed human hepatocyte growth factor (hHGF) in primary rat DPSCs and evaluated the effects of HGF overexpression on the biological behaviors and therapeutic efficacy of grafted DPSCs in cirrhosis. Liver cirrhosis was induced via the intraperitoneal injection of CCl4 twice weekly for 12 weeks and was verified through histopathological and serological assays. HGF was overexpressed in DPSCs via transduction with a hHGF-lentiviral vector and confirmed based on the elevated expression and secretion of HGF. The HGF-overexpressing DPSCs were transplanted into rats intravenously. The HGF-overexpressing DPSCs showed increased survival and hepatogenic differentiation in host liver tissue at 6 weeks after grafting. They also exhibited a significantly greater repair potential in relation to cirrhosis pathology and impaired liver function than did DPSCs expressing HGF at physiological levels. Our study may provide an experimental basis for the development of novel methods for the treatment of liver cirrhosis in clinical practice.

Comparison of bone regeneration in alveolar bone of dogs on mineralized collagen grafts with two composition ratios of nano-hydroxyapatite and collagen.

To study the effect of two composition ratios of nano-hydroxyapatite and collagen (NHAC) composites on repairing alveolar bone defect of dogs. Eighteen healthy adult dogs were randomly divided into three groups. Two kinds of the NHAC composites were prepared according to the constituent ratios of 3:7 and 5:5; immediately after extraction of the mandibular second premolars, each kind of the NHAC composite was implanted into extraction socket, respectively: Group I, nHA/Col = 3:7; Group II, nHA/Col = 5:5 and Group III, blank control group. The bone-repairing ability of the two grafts was separately analyzed by morphometric measurement, X-ray tomography examination and biomechanical analysis at 1st, 3rd and 6th month post-surgical, respectively. The NHAC composites were absorbed gradually after implanting into alveolar bone defect and were replaced by new bone. The ratios of new bone formation of Group I was significantly higher than that of Group II after 3 months (P < 0.05). The structure and bioactive performance can be improved when the ratio between the collagen and the hydroxyapatite was reasonable, and the repairing ability and effect in extraction sockets are obviously better.

Clinical evaluations of mineralized collagen in the extraction sites preservation.

The purpose of this study was to explore the different effects between biomimetic mineralized collagen (MC) and ordinary physically blended hydroxyapatite/collagen (HA/Col) composite in evaluating new bone formation and regenerated bone height in human extraction sockets. Thirty-four patients who cannot retain teeth caused by trauma or decay were randomly selected from Department of Stomatology of Dongzhimen Hospital from December 2013 to December 2014. The patients were randomly divided into two groups. After the operation of tooth extraction, 17 patients were implanted with biomimetic MC (MC group), and other 17 patients were implanted with ordinary physically blended nHA/Col composite (nHA/Col group). X-ray positioning projection by auto-photographing was taken to test the distance between the lowest position and the neighboring CEJm-CEJd immediately, 1 month and 3 months after the operation. The height of new bone formation of the MC group was significantly higher than the nHA/Col group. Biomimetic MC showed better clinical outcomes in the bone formation for extraction site preservation and would have broad application prospect in the field of oral and maxillofacial surgeries.

Self­assembled monolayers of alkanethiolates on surface chemistry groups in osteosarcoma cells.

Cell biomedical behavior is influenced by a number of factors, and the extracellular matrix (ECM) of the cellular microenvironment affects certain cancer cells. In the current study, U­2OS cells were cultured on gold surfaces modified with different terminal chemical groups [methyl (­CH3), amino (­NH2), hydroxyl (­OH) and carboxyl (­COOH)]. The results revealed that different chemical surfaces convey different behaviors. The density of the different functional surfaces was confirmed by atomic force microscopy. Cell morphology, proliferation rate and cell cycle were investigated using scanning electron microscopy, cell counting and flow cytometry. In conclusion, the type of chemical group on a biomaterial is an important property for the growth of osteosarcoma cells; ­NH2 and ­COOH surfaces sustained visible cell adhesion and promoted cell growth.

Biological evaluation of human degenerated nucleus pulposus cells in functionalized self-assembling peptide nanofiber hydrogel scaffold.

Nucleus pulposus (NP) tissue engineering has been proposed as a novel biological treatment for early-stage intervertebral disc degeneration. In this study, a novel functional self-assembling peptide PKP was first designed by linking the short functional motif of bone morphogenetic protein-7 (BMP7) to the C-terminal of RADA16-I, and another new functional self-assembling peptide was obtained by mixing RKP with RADA16-I. Then, the biocompatibilities and bioactivities of RKP and RAD-RKP for human degenerated nucleus pulposus cells (hNPCs) were studied in vitro. Atomic force microscopy and scanning electron microscopy (SEM) confirmed that both RKP and RAD-RKP could self-assemble into three-dimensional (3D) nanofiber hydrogel scaffolds in a culture medium at 37°C. After the hNPCs were cultured in 3D scaffolds, both RKP and RAD-RKP exhibited reliable attachment and extremely low cytotoxicities (<14%), which were verified by SEM and cytotoxity assays, respectively. Our results also showed that the functional-based scaffolds could increase the proliferation and migration of hNPCs after 7 days compared with culture plates and pure RADA16-I. Quantitative real-time polymerase chain reaction demonstrated that the expressions of collagen II α1, Sox-9, and aggrecan were upregulated, while collagen I α1 was downregulated by functional-based scaffolds after 28 days. Furthermore, we also confirmed that RAD-RKP exhibited a higher hNPC proliferation, migration, and expression of Sox-9 and aggrecan compared with pure RKP. Therefore, the results of this study indicated that the BMP7 short motif-designed functional self-assembling peptide nanofiber hydrogels could be used as excellent scaffolds in NP tissue engineering, and RAD-RKP might have further potential application in human mild degenerated NP tissue regeneration.

Activation of the ERK1/2 signaling pathway during the osteogenic differentiation of mesenchymal stem cells cultured on substrates modified with various chemical groups.

The current study examined the influence of culture substrates modified with the functional groups -OH, -COOH, -NH2, and -CH3 using SAMs technology, in conjunction with TAAB control, on the osteogenic differentiation of rabbit BMSCs. The CCK-8 assay revealed that BMSCs exhibited substrate-dependent cell viability. The cells plated on -NH2- and -OH-modified substrates were well spread and homogeneous, but those on the -COOH- and -CH3-modified substrates showed more rounded phenotype. The mRNA expression of BMSCs revealed that -NH2-modified substrate promoted the mRNA expression and osteogenic differentiation of the BMSCs. The contribution of ERK1/2 signaling pathway to the osteogenic differentiation of BMSCs cultured on the -NH2-modified substrate was investigated in vitro. The -NH2-modified substrate promoted the expression of integrins; the activation of FAK and ERK1/2. Inhibition of ERK1/2 activation by PD98059, a specific inhibitor of the ERK signaling pathway, blocked ERK1/2 activation in a dose-dependent manner, as revealed for expression of Cbf α -1 and ALP. Blockade of ERK1/2 phosphorylation in BMSCs by PD98059 suppressed osteogenic differentiation on chemical surfaces. These findings indicate a potential role for ERK in the osteogenic differentiation of BMSCs on surfaces modified by specific chemical functional groups, indicating that the microenvironment affects the differentiation of BMSCs. This observation has important implications for bone tissue engineering.

Enamel distribution, structure and mechanical alterations in col1-caPPR mice molar.

Enamel mineralisation is a highly controlled process in which the deposition, growth, and maturation of inorganic crystallites are regulated by secreted matrix proteins at the molecular and cellular level. Maxillary and mandibular first molars from the col1-caPPR mutants as well as normal controls aged for 12 weeks were observed by SEM and nanoindentation, respectively. Several types of aberrations in enamel distribution and crystal organisation were encountered in the transgenic molars. Also, the gene alteration resulted in degradation by as much as 23.42% in hardness and 17.56% in the elastic modulus. These data suggested that gene mutation altered the ameloblastic differentiation and movement, resulting in variations of crystal arrangement patterns, aberrations of enamel distribution, and degradation of mechanical behaviour. Furthermore, the col1-caPPR mouse model was determined as useful for studying how the genes modulate the biomineralisation process.

Injectable bone cement based on mineralized collagen.

A novel injectable bone cement based on mineralized collagen was reported in this paper. The cement was fabricated by introducing calcium sulfate hemihydrate (CaSO(4).1/2H(2)O, CSH) into nano-hydroxyapatite/collagen (nHAC). The workability, in vitro degradation, in vitro and in vivo biocompatibility of the cement (nHAC/CSH) were studied. The comparative tests via in vitro and in vivo showed that the nHAC/CSH composite cement processed better biocompatibiltiy than that of pure CSH cement. The results implied that this new injectable cement should be very promising for bone repair.

The enamel softening and loss during early erosion studied by AFM, SEM and nanoindentation.

Enamel dissolution occurs when it contacts with acids produced by plaque bacteria, foods or drinks. There have been numerous and varied studies quantifying and characterizing the rate, extent and chemical aspects of enamel erosion; however, there is still hot debate about the amounts of enamel softening and loss. The objective of this study was to measure the enamel erosion process with high accuracy. Native third molars were partly eroded in citric acid (pH = 3.8) with some domains protected by a Ti coating layer. The surface morphology and structures before and after exposure to citric acid for different time periods were studied by AFM, SEM and nanoindentation, respectively. Based on this, the functions between the amounts of enamel softening, loss and erosion time were established for the first time. It was demonstrated that AFM, SEM and nanoindentation were suitable for measuring the early stages of enamel demineralization qualitatively and quantitatively.