Differential effect of water-soluble chitin on collagen synthesis of human bone marrow stem cells and human periodontal ligament stem cells.

Human bone marrow stem cells (hBMSCs) represent a promising regenerative material because of their mutipotency, including their ability to regenerate collagenous soft tissues. We previously found that water-soluble chitin (WSC) enhances the ability of human periodontal ligament stem cells (hPDLSCs) to synthesize collagen tissue. The aim of this study was to determine the effects of WSC on hBMSCs and hPDLSCs for the collagen synthesis both in vitro and in vivo. hBMSCs and hPDLSCs were isolated and expanded with or without 0.3 mg/mL WSC. A series of in vitro and in vivo analyses were performed to evaluate their characteristics as stem cell populations. Then, collagen and hydroxyproline assays were conducted using both in vitro and in vivo assay models, and the real-time polymerase chain reaction was performed to analyze the expression of collagen-related markers. WSC-treated and nontreated hBMSCs and hPDLSCs were transplanted into immunocompromised mice, and histology and immunohistochemistry analyses were conducted after 8 weeks. The in vitro results showed that those cells possessed the characteristics of mesenchymal stem cells. The amount of soluble collagen synthesized was significantly greater in WSC-treated hBMSCs than in the nontreated group; conversely, treatment of hPDLSCs with WSC decreased the formation of soluble collagen. The amount of insoluble collagen synthesized was greater in the WSC-treated groups than in the nontreated groups for both hBMSCs and hPDLSCs. The hydroxyproline contents of the regenerated soluble and insoluble collagens were similar. The expressions of mRNA for collagen types I-V, hyaluronic acid synthase 1 (HAS1), HAS2, and HAS3, and the LOX family were higher in WSC-treated hPDLSCs than in the nontreated group, whereas WSC increased the expression of collagen type III and decreased that of collagen type I in hBMSCs. The histology and immunohistochemistry results revealed that WSC significantly increased the amount of collagen formed in vivo by both types of stem cells. Collectively, treatment with WSC significantly enhanced the collagen-forming potentials of hBMSCs and hPDLSCs, but the collagen they produced exhibited distinctively different characteristics. These findings suggest that the appropriate stem-cell source should be chosen based on the purpose of the required regenerated tissue.

Anti-allergic activity of crystallinity controlled N-acetyl glucosamine.

CONTEXT: Chitin is the polysaccharide and is found in insects, parasites and fungi. Chitin has shown various immunological effects in in vivo and in vitro models. In this study, crystallinity controlled N-acetyl glucosamine (CCG) which has a high solubility was prepared from the low molecular weight chitosan. However, the effect of CCG in an allergic response is not clear. OBJECTIVE: To investigate the effect and regulatory mechanism of CCG on allergic responses. METHODS: To demonstrate the effect of CCG, we induced systemic anaphylactic shock, ear swelling response, passive cutaneous anaphylaxis (PCA), and inflammatory reaction. RESULTS: CCG inhibited the compound 48/80-induced systemic anaphylactic shock and ear swelling responses. IgE-mediated PCA was inhibited by the oral administration or topical application of CCG. Histamine and ß-hexosaminidase release from mast cells was decreased with the treatment of CCG. CCG also inhibited phorbol 12-myristate 13-acetate and calcium ionophore A23187-induced interleukin-1ß production and mRNA expression by suppressing NF-κB activation and IκBα phosphorylation. Furthermore, CCG suppressed the activation of caspase-1. CONCLUSION: These results suggest that CCG may have beneficial applicability as a candidate for an anti-allergic agent.

Novel application of human periodontal ligament stem cells and water-soluble chitin for collagen tissue regeneration: in vitro and in vivo investigations.

Human periodontal ligament stem cells (hPDLSCs) have been proposed as an alternative to conventional cosmetic fillers because they display an innate ability to synthesize collagen. The aims of this study were to determine the effects of water-soluble chitin (WSC) on the proliferation and migration of hPDLSCs, and to quantify collagen synthesis in vitro and in vivo compared with human adipose-derived stem cell (hADSC)s. hPDLSCs were isolated from healthy extracted teeth, and the cell proliferation and cell migration capacities of untreated hPDLSCs (control group) and WSC-treated hPDLSCs (test group) were compared. Insoluble/soluble collagen synthesis were also assessed, and collagen related markers were evaluated including lysyl oxidase (LOX), lysyl oxidase like (LOXL)1, LOXL2, and hydroxyproline. In vivo collagen formation was examined by transplanting hyaluronic acid as a cell carrier into the subcutaneous pockets of immunocompromised mice in the control and test groups; histology and immunohistochemistry analyses were performed 4 (n=4) and 8 (n=4) weeks later. There was a dose-dependent enhancement of hPDLSCs proliferation in the test group, and a concomitant reduction in cell migration. The amount of insoluble collagen formed was greater in the test group than in the control group (p<0.05), whereas soluble collagen formation was significantly reduced in the test group (p<0.05). The histology and immunohistochemistry results revealed that the amount of collagen formed in vivo was greater in WSC-treated hPDLSCs than in the control cells at 4 and 8 weeks (p<0.05), and histometric analysis at 8 weeks revealed that enhancement of collagen formation by hPDLSCs was greater than by hADSCs. These results indicate that WSC modulates the properties of hPDLSCs, rendering them more suitable for cosmetic soft-tissue augmentation.

Effects of chitosan nonwoven membrane on periodontal healing of surgically created one-wall intrabony defects in beagle dogs.

The purpose of this study was to investigate the periodontal regenerative effects of a chitosan nonwoven membrane applied to surgically created preclinical one-wall intrabony defects in beagle dogs. One-wall intrabony defects (4 x 4 x 4 mm) were surgically created bilaterally in the mandibular second and fourth premolars of six beagle dogs. The surgical control group received a flap operation only. The resorbable-membrane (RM) group was treated with resorbable membrane. The chitosan-nonwoven-membrane (CNWM) group was treated with chitosan nonwoven membrane. The amount of junctional epithelium migration and the amount of connective tissue adhesion did not show any statistically significant differences among the groups. However, the amount of suprabony cementum regeneration, intrabony cementum regeneration, and alveolar bone regeneration showed significant differences (p < 0.05) between CNWM site group and control group. The results demonstrate the regenerative effects of the chitosan nonwoven membrane in one-wall intrabony defects of beagle dogs. The chitosan nonwoven membrane has the potential to support the cementum and bone regeneration, possibly by providing the conditions needed for guided tissue regeneration in the one-wall intrabony periodontal defects of beagle dogs.