Jiaqiang Liu

Effect of nano-structured bioceramic surface on osteogenic differentiation of adipose derived stem cells

Tissue engineering strategies to construct vascularized bone grafts potentially revolutionize the treatment of massive bone loss. The surface topography of the grafts plays critical roles on bone regeneration, while adipose derived stem cells (ASCs) are known for their capability to promote osteogenesis and angiogenesis when applied to bone defects. In the present study, the effects of hydroxyapatite (HAp) bioceramic scaffolds with nanosheet, nanorod, and micro-nano-hybrid (the hybrid of nanorod and microrod) surface topographies on attachment, proliferation and osteogenic differentiation, as well as the expression of angiogenic factors of rat ASCs were systematically investigated. The results showed that the HAp bioceramic scaffolds with the micro-/nano-topography surfaces significantly enhanced cell attachment and viability, alkaline phosphatase (ALP) activity, and mRNA expression levels of osteogenic markers and angiogenic factors of ASCs. More importantly, the biomimetic feature of the hierarchical micro-nano-hybrid surface topography showed the highest stimulatory effect. The activation in Akt signaling pathway was observed in ASCs cultured on HAp bioceramics with nanorod, and micro-nano-hybrid surface topographies. Moreover, these induction effects could be repressed by Akt signaling pathway inhibitor LY294002. Finally, the in vivo bone regeneration results of rat critical-sized calvarial defect models confirmed that the combination of the micro-nano-hybrid surface and ASCs could significantly enhance both osteogenesis and angiogenesis as compared with the control HAp bioceramic scaffold with traditional smooth surface. Our results suggest that HAp bioceramic scaffolds with micro-nano-hybrid surface can act as cell carrier for ASCs, and consequently combine with ASCs to construct vascularized tissue-engineered bone.

Akermanite bioceramics promote osteogenesis, angiogenesis and suppress osteoclastogenesis for osteoporotic bone regeneration.

It is a big challenge for bone healing under osteoporotic pathological condition with impaired angiogenesis, osteogenesis and remodeling. In the present study, the effect of Ca, Mg, Si containing akermanite bioceramics (Ca2MgSi2O7) extract on cell proliferation, osteogenic differentiation and angiogenic factor expression of BMSCs derived from ovariectomized rats (BMSCs-OVX) as well as the expression of osteoclastogenic factors was evaluated. The results showed that akermanite could enhance cell proliferation, ALP activity, expression of Runx2, BMP-2, BSP, OPN, OCN, OPG and angiogenic factors including VEGF and ANG-1. Meanwhile, akermanite could repress expression of osteoclastogenic factors including RANKL and TNF-α. Moreover, akermanite could activate ERK, P38, AKT and STAT3 signaling pathways, while crosstalk among these signaling pathways was evident. More importantly, the effect of akermanite extract on RANKL-induced osteoclastogenesis was evaluated by TRAP staining and real-time PCR assay. The results showed that akermanite could suppress osteoclast formation and expression of TRAP, cathepsin K and NFATc1. The in vivo experiments revealed that akermanite bioceramics dramatically stimulated osteogenesis and angiogenesis in an OVX rat critical-sized calvarial defect model. All these results suggest that akermanite bioceramics with the effects of Mg and Si ions on osteogenesis, angiogenesis and osteoclastogenesis are promising biomaterials for osteoporotic bone regeneration.

Designing ordered micropatterned hydroxyapatite bioceramics to promote the growth and osteogenic differentiation of bone marrow stromal cells

Patterned structured surfaces are very useful to control a cells microenvironment and to modulate certain cellular responses, such as cell adhesion, migration, proliferation, and differentiation. Herein, ordered micropatterns constructed by a quadrate convex with different sizes were fabricated on a hydroxyapatite [Ca10(PO4)6(OH)2, HAp] bioceramic surface using an ordered micropatterned nylon sieve as templates. The height, width and space of the convex for the patterns could be facilely regulated via simply tailoring the meshes of the template. Compared with traditional samples with flat surfaces, the fabricated HAp bioceramics with micropatterned surfaces possessed better wettability and higher surface energy, which significantly promoted the adhesion, proliferation, and osteogenic differentiation of rat bone marrow stromal cells (bMSCs). Furthermore, using a pattern size close to that of the cell size showed a better stimulation of cell response compared with larger pattern sizes. Our study suggests that the fabrication of micropatterned structured HAp bioceramics is critical for designing optimal biomaterials for bone regeneration and cell culture substrate applications.

The synergistic effects of Sr and Si bioactive ions on osteogenesis, osteoclastogenesis and angiogenesis for osteoporotic bone regeneration

Bioactive ions released from bioceramics play important roles in bone regeneration; however, it is unclear how each ionic composition in complex bioceramics exerts its specific effect on bone regeneration. The aim of this study is to elucidate the functional effects of Sr and Si ions in bioceramics on the regeneration of osteoporotic bone. A model bioceramic with Sr- and Si-containing components (SMS) was successfully fabricated and the effects of ionic products from SMS bioceramics on the osteogenic, osteoclastic and angiogenic differentiation of rBMSCs-OVX and RANKL-induced osteoclasts were investigated. The results showed that SMS bioceramics could enhance ALP activity and expression of Col 1, OCN, Runx2, and angiogenic factors including VEGF and Ang-1. SMS bioceramics not only rebalanced the OPG/RANKL ratio of rBMSCs-OVX at early stage, but also repressed RANKL-induced osteoclast formation and expression of TRAP, DC-STAMP, V-ATPase a3, and NFATc1. The synergistic effects of Sr and Si ions were further investigated as compared with those of similar concentrations of Sr and Si ions alone. Sr and Si ions possessed synergistic effects on osteogenesis, osteoclastogenesis, and angiogenesis, attributed to the dominant effects of Sr ions on enhancing angiogenesis and repressing osteoclastogenesis, and the dominant effects of Si ions on stimulating osteogenesis. The in vivo study using critical-size mandibular defects of OVX rat models showed that SMS bioceramics could significantly enhance bone formation and mineralization compared with β-TCP bioceramics. Our results are the first to elucidate the specific effect of each ion from bioceramics on osteogenesis, osteoclastogenesis and angiogenesis for osteoporotic bone regeneration, paving the way for the design of functional biomaterials with complex compositions for tissue engineering and regenerative medicine. STATEMENT OF SIGNIFICANCE Bioactive ions released from bioceramics play important roles for bone regeneration; however, it is unclear how each of ionic compositions in complex bioceramics exerts its specific effect on bone regeneration. The aim of present study is to elucidate the functional effects of Sr and Si ions in complex bioceramics on the regeneration of osteoporotic bone. A model bioceramic with Sr and Si-containing components (SMS) was successfully fabricated and the effects of ionic products from SMS bioceramics on the osteogenic, osteoclastic and angiogenic differentiation of rBMSCs-OVX and RANKL-induced osteoclasts were investigated. The results showed that SMS bioceramics could enhance ALP activity and expression of Col 1, OCN, Runx2 and angiogenic factors including VEGF and Ang-1. SMS bioceramics not only rebalanced the ratio of OPG/RANKL of OVX-BMSCs at early stage, but also repressed RANKL-induced osteoclast formation and expression of TRAP, DC-STAMP, V-ATPase a3, and NFATc1. The synergistic effects of Sr and Si ions were further investigated as compared with the similar concentration of Sr and Si ions alone. It was found that Sr and Si ions possessed synergistic effects on osteogenesis, osteoclastogenesis and angiogenesis, attributed to the dominant effects of Sr ions on enhancing angiogenesis and repressing osteoclastogenesis, and the dominant effects of Si ions on stimulating osteogenesis. The in vivo study using critical-size mandibular defects of OVX rat models showed that SMS bioceramics could significantly enhance bone formation and mineralization as compared with β-TCP bioceramics. It is suggested that SMS bioceramics may be a promising biomaterial for osteoporotic bone regeneration. To our knowledge, this is the first time to elucidate the specific effect of each ion from bioceramics on osteogenesis, osteoclastogenesis and angiogenesis for osteoporotic bone regeneration, paving the way to design functional biomaterials with complex compositions for tissue engineering and regenerative medicine.

The synergetic effect of nano-structures and silicon-substitution on the properties of hydroxyapatite scaffolds for bone regeneration

Control over the morphology and chemical composition of hydroxyapatite (HAp) bioceramic scaffolds is of great importance for their applications. In the present study, Si-substituted HAp bioceramic scaffolds with controllable morphologies (nanosheets and nanorods) were fabricated via hydrothermal treatment of calcium silicate scaffolds as precursors in NaH2PO4 and Na3PO4 aqueous solutions, respectively. Moreover, the effects of surface morphologies and Si substitution on cell attachment, proliferation, and osteogenic differentiation of rat bone marrow stromal cells (rBMSCs) were systematically investigated in vitro. The results showed that nano-topography surfaces could enhance cell attachment, cell proliferation, alkaline phosphatase (ALP) activity, and mRNA expression levels of collagen 1 (COL1), bone morphogenetic protein 2 (BMP-2), bone sialoprotein (BSP) and osteopontin (OPN). Moreover, the Si substitution could further promote cell proliferation and osteogenic differentiation, while Si-substituted bioceramics with a nanorod surface possessed the highest stimulatory effect. More importantly, the in vivo rat critical-sized calvarial defect model confirmed that HAp bioceramic scaffolds with nanosheet and nanorod surfaces showed definitive bone regeneration as compared with control HAp bioceramic scaffolds with a traditional smooth surface. Moreover, Si substitution could synergistically enhance bone regeneration and mineralization, while Si-substituted HAp bioceramic scaffolds with a nanorod surface achieved the best bone repair ability. The present study suggests that the modification of the surface morphology and Si substitution on the HAp bioceramic scaffold may be an effective synergistic strategy to improve its clinical performance.

Effect of micro-nano-hybrid structured hydroxyapatite bioceramics on osteogenic and cementogenic differentiation of human periodontal ligament stem cell via Wnt signaling pathway

The surface structure of bioceramic scaffolds is crucial for its bioactivity and osteoinductive ability, and in recent years, human periodontal ligament stem cells have been certified to possess high osteogenic and cementogenic differential ability. In the present study, hydroxyapatite (HA) bioceramics with micro-nano-hybrid surface (mnHA [the hybrid of nanorods and microrods]) were fabricated via hydrothermal reaction of the α-tricalcium phosphate granules as precursors in aqueous solution, and the effects of mnHA on the attachment, proliferation, osteogenic and cementogenic differentiations of human periodontal ligament stem cells as well as the related mechanisms were systematically investigated. The results showed that mnHA bioceramics could promote cell adhesion, proliferation, alkaline phosphatase (ALP) activity, and expression of osteogenic/cementogenic-related markers including runt-related transcription factor 2 (Runx2), ALP, osteocalcin (OCN), cementum attachment protein (CAP), and cementum protein (CEMP) as compared to the HA bioceramics with flat and dense surface. Moreover, mnHA bioceramics stimulated gene expression of low-density lipoprotein receptor-related protein 5 (LRP5) and β-catenin, which are the key genes of canonical Wnt signaling. Moreover, the stimulatory effect on ALP activity and osteogenic and cementogenic gene expression, including that of ALP, OCN, CAP, CEMP, and Runx2 of mnHA bioceramics could be repressed by canonical Wnt signaling inhibitor dickkopf1 (Dkk1). The results suggested that the HA bioceramics with mnHA could act as promising grafts for periodontal tissue regeneration.

Three-dimensional analysis of soft tissue changes in full-face view after surgical correction of skeletal class III malocclusion.

ObjectiveThe objective of this study was to evaluate changes in soft tissue in full-face view because of surgical correction of skeletal Class III malocclusion, using 3-dimensional (3D) laser scanning. MethodsTwenty-seven subjects with skeletal Class III malocclusion [11 males; mean age (SD), 24.0 (5.7) years] underwent bilateral sagittal split ramus osteotomy for mandibular setback combined with Lefort I osteotomy with/without maxillary advancement. Twelve patients (group 1) had mandibular setback surgery, and the other 15 (group 2) had combination surgery. Lateral cephalograms and 3D facial scan images were assessed preoperatively and postoperatively. The facial widths upon superimposition of 3D facial images were measured in the same coordinates using a Rapidform 2006 system. Paired and independent t tests were done for statistical analysis. ResultsThe midface soft tissue broadened significantly above the cheilion plane postoperatively (P < 0.05). A larger change was observed nearer to subnasale plane, and a similar trend was seen among the horizontal planes in 1- or 2-jaw surgery groups. The widths from the exocanthion plane to the subnasale plane increased more in group 2 [mean (SD), 4.45 (2.45) mm, 8.71 (2.92) mm, and 7.62 (3.13) mm] than those in group 1 [mean (SD), 1.26 (0.97) mm, 1.84 (1.06) mm, and 1.35 (0.65) mm], and this difference was significant (P < 0.05). There was a decrease below the cheilion plane with mandibular setback between groups, but this difference was not significant. ConclusionsThe measurement method used here for the shape outline of the lateral parts of the face could provide quantitative data for the clinical evaluation and objective analysis of the human face in full-face view. The midface soft tissue in subjects with skeletal Class III malocclusion exhibited a greater increase in width after bimaxillary surgery procedures than mandibular setback-only surgery.

ADAM10 is essential for cranial neural crest-derived maxillofacial bone development.

Growth disorders of the craniofacial bones may lead to craniofacial deformities. The majority of maxillofacial bones are derived from cranial neural crest cells via intramembranous bone formation. Any interruption of the craniofacial skeleton development process might lead to craniofacial malformation. A disintegrin and metalloprotease (ADAM)10 plays an essential role in organ development and tissue integrity in different organs. However, little is known about its function in craniofacial bone formation. Therefore, we investigated the role of ADAM10 in the developing craniofacial skeleton, particularly during typical mandibular bone development. First, we showed that ADAM10 was expressed in a specific area of the craniofacial bone and that the expression pattern dynamically changed during normal mouse craniofacial development. Then, we crossed wnt1-cre transgenic mice with adam10-flox mice to generate ADAM10 conditional knockout mice. The stereomicroscopic, radiographic, and von Kossa staining results showed that conditional knockout of ADAM10 in cranial neural crest cells led to embryonic death, craniofacial dysmorphia and bone defects. Furthermore, we demonstrated that impaired mineralization could be triggered by decreased osteoblast differentiation, increased cell death. Overall, these findings show that ADAM10 plays an essential role in craniofacial bone development.