Hydrogels/therapeutic use* ; Tissue Engineering ; Gelatin ; Intervertebral Disc/surgery* ; Methacrylates ; Tissue Scaffolds

Pulp loss is accompanied by the functional impairment of defense, sensory, and nutrition supply. The approach based on endogenous stem cells is a potential strategy for pulp regeneration. However, endogenous stem cell sources, exogenous regenerative signals, and neovascularization are major difficulties for pulp regeneration based on endogenous stem cells. Therefore, the purpose of our research is to seek an effective cytokines delivery strategy and bioactive materials to reestablish an ideal regenerative microenvironment for pulp regeneration. In in vitro study, we investigated the effects of Wnt3a, transforming growth factor-beta 1, and bone morphogenetic protein 7 (BMP7) on human dental pulp stem cells (h-DPSCs) and human umbilical vein endothelial cells. 2D and 3D culture systems based on collagen gel, matrigel, and gelatin methacryloyl were fabricated to evaluate the morphology and viability of h-DPSCs. In in vivo study, an ectopic nude mouse model and an in situ beagle dog model were established to investigate the possibility of pulp regeneration by implanting collagen gel loading BMP7. We concluded that BMP7 promoted the migration and odontogenic differentiation of h-DPSCs and vessel formation. Collagen gel maintained the cell adhesion, cell spreading, and cell viability of h-DPSCs in 2D or 3D culture. The transplantation of collagen gel loading BMP7 induced vascularized pulp-like tissue regeneration in vivo. The injectable approach based on collagen gel loading BMP7 might exert promising therapeutic application in endogenous pulp regeneration.
Animals ; Bone Morphogenetic Protein 7/pharmacology* ; Cell Differentiation ; Cells, Cultured ; Collagen/pharmacology* ; Dental Pulp ; Dogs ; Endothelial Cells ; Gelatin ; Humans ; Methacrylates ; Mice ; Regeneration ; Stem Cells

Objective: To prepare graphene oxide (GO)-containing gelatin methacrylate anhydride (GelMA) hydrogel and to investigate the effects of in situ photopolymerized GO-GelMA composite hydrogel in wound vascularization of full-thickness skin defect in mice. Methods: The experimental study method was used. The 50 μL of 0.2 mg/mL GO solution was evenly applied onto the conductive gel, and the structure and size of GO were observed under field emission scanning electron microscope after drying. Human skin fibroblasts (HSFs) were divided into 0 μg/mL GO (without GO solution, the same as below) group, 0.1 μg/mL GO group, 1.0 μg/mL GO group, 5.0 μg/mL GO group, and 10.0 μg/mL GO group treated with GO of the corresponding final mass concentration, and the absorbance value was detected using a microplate analyzer after 48 h of culture to reflect the proliferation activity of cells (n=6). HSFs and human umbilical vein vascular endothelial cells (HUVECs) were divided into 0 μg/mL GO group, 0.1 μg/mL GO group, 1.0 μg/mL GO group, and 5.0 μg/mL GO group treated with GO of the corresponding final mass concentration, and the migration rates of HSFs at 24 and 36 h after scratching (n=5) and HUVECs at 12 h after scratching (n=3) were detected by scratch test, and the level of vascular endothelial growth factor (VEGF) secreted by HSFs after 4, 6, and 8 h of culture was detected by enzyme-linked immunosorbent assay method (n=3). The prepared GO-GelMA composite hydrogels containing GO of the corresponding final mass concentration were set as 0 μg/mL GO composite hydrogel group, 0.1 μg/mL GO composite hydrogel group, 1.0 μg/mL GO composite hydrogel group, and 5.0 μg/mL GO composite hydrogel group to observe their properties before and after cross-linking, and to detect the release of GO after soaking with phosphate buffer solution for 3 and 7 d (n=3). The full-thickness skin defect wounds were made on the back of 16 6-week-old female C57BL/6 mice. The mice treated with in situ cross-linked GO-GelMA composite hydrogel containing GO of the corresponding final mass concentration were divided into 0 μg/mL GO composite hydrogel group, 0.1 μg/mL GO composite hydrogel group, 1.0 μg/mL GO composite hydrogel group, and 5.0 μg/mL GO composite hydrogel group according to the random number table, with 4 mice in each group. The general condition of wound was observed and the wound healing rate was calculated on 3, 7, and 14 d of treatment, the wound blood perfusion was detected by laser Doppler flowmetry on 3, 7, and 14 d of treatment and the mean perfusion unit (MPU) ratio was calculated, and the wound vascularization on 7 d of treatment was observed after hematoxylin-eosin staining and the vascular density was calculated (n=3). The wound tissue of mice in 0 μg/mL GO composite hydrogel group and 0.1 μg/mL GO composite hydrogel group on 7 d of treatment was collected to observe the relationship between the distribution of GO and neovascularization by hematoxylin-eosin staining (n=3) and the expression of VEGF by immunohistochemical staining. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and Tukey's method. Results: GO had a multilayered lamellar structure with the width of about 20 μm and the length of about 50 μm. The absorbance value of HSFs in 10.0 μg/mL GO group was significantly lower than that in 0 μg/mL GO group after 48 h of culture (q=7.64, P<0.01). At 24 h after scratching, the migration rates of HSFs were similar in the four groups (P>0.05); at 36 h after scratching, the migration rate of HSFs in 0.1 μg/mL GO group was significantly higher than that in 0 μg/mL GO group, 1.0 μg/mL GO group, and 5.0 μg/mL GO group (with q values of 7.48, 10.81, and 10.20, respectively, P<0.01). At 12 h after scratching, the migration rate of HUVECs in 0.1 μg/mL GO group was significantly higher than that in 0 μg/mL GO group, 1.0 μg/mL GO group, and 5.0 μg/mL GO group (with q values of 7.11, 8.99, and 14.92, respectively, P<0.01), and the migration rate of HUVECs in 5.0 μg/mL GO group was significantly lower than that in 0 μg/mL GO group and 1.0 μg/mL GO group (with q values of 7.81 and 5.33, respectively, P<0.05 or P<0.01 ). At 4 and 6 h of culture, the VEGF expressions of HSFs in the four groups were similar (P>0.05); at 8 h of culture, the VEGF expression of HSFs in 0.1 μg/mL GO group was significantly higher than that in 0 μg/mL GO group and 5.0 μg/mL GO group (with q values of 4.75 and 4.48, respectively, P<0.05). The GO-GelMA composite hydrogels in the four groups were all red liquid before cross-linking, which turned to light yellow gel after cross-linking, with no significant difference in fluidity. The GO in the GO-GelMA composite hydrogel of 0 μg/mL GO composite hydrogel group had no release of GO at all time points; the GO in the GO-GelMA composite hydrogels of the other 3 groups was partially released on 3 d of soaking, and all the GO was released on 7 d of soaking. From 3 to 14 d of treatment, the wounds of mice in the 4 groups were covered with hydrogel dressings, kept moist, and gradually healed. On 3, 7, and 14 d of treatment, the wound healing rates of mice in the four groups were similar (P>0.05). On 3 d of treatment, the MPU ratio of wound of mice in 0.1 μg/mL GO composite hydrogel group was significantly higher than that in 0 μg/mL GO composite hydrogel group, 1.0 μg/mL GO composite hydrogel group, and 5.0 μg/mL GO composite hydrogel group (with q values of 10.70, 11.83, and 10.65, respectively, P<0.05 or P<0.01). On 7 and 14 d of treatment, the MPU ratios of wound of mice in the four groups were similar (P>0.05). The MPU ratio of wound of mice in 0.1 μg/mL GO composite hydrogel group on 7 d of treatment was significantly lower than that on 3 d of treatment (q=14.38, P<0.05), and that on 14 d of treatment was significantly lower than that on 7 d of treatment (q=27.78, P<0.01). On 7 d of treatment, the neovascular density of wound of mice on 7 d of treatment was 120.7±4.1 per 200 times of visual field, which was significantly higher than 61.7±1.3, 77.7±10.2, and 99.0±7.9 per 200 times of visual field in 0 μg/mL GO composite hydrogel group, 1.0 μg/mL GO composite hydrogel group, and 5.0 μg/mL GO composite hydrogel group (with q values of 12.88, 7.79, and 6.70, respectively, P<0.01), and the neovascular density of wound of mice in 1.0 μg/mL GO composite hydrogel group and 5.0 μg/mL GO composite hydrogel group was significantly higher than that in 0 μg/mL GO composite hydrogel group (with q values of 5.10 and 6.19, respectively, P<0.05). On 7 d of treatment, cluster of new blood vessels in wound of mice in 0.1 μg/mL GO composite hydrogel group was significantly more than that in 0 μg/mL GO composite hydrogel group, and the new blood vessels were clustered near the GO; a large amount of VEGF was expressed in wound of mice in 0.1 μg/mL GO composite hydrogel group in the distribution area of GO and new blood vessels. Conclusions: GO with mass concentration lower than 10.0 μg/mL had no adverse effect on proliferation activity of HSFs, and GO of 0.1 μg/mL can promote the migration of HSFs and HUVECs, and can promote the secretion of VEGF in HSFs. In situ photopolymerized of GO-GelMA composite hydrogel dressing can promote the wound neovascularization of full-thickness skin defect in mice and increase wound blood perfusion in the early stage, with GO showing an enrichment effect on angiogenesis, and the mechanism may be related to the role of GO in promoting the secretion of VEGF by wound cells.
Anhydrides ; Animals ; Endothelial Cells ; Eosine Yellowish-(YS) ; Female ; Gelatin/pharmacology* ; Graphite ; Hematoxylin ; Humans ; Hydrogels/pharmacology* ; Methacrylates ; Mice ; Mice, Inbred C57BL ; Neovascularization, Pathologic ; Skin Abnormalities ; Vascular Endothelial Growth Factor A

Wound repair is a common clinical problem, which seriously affects the quality of life of patients and also brings a heavy burden to the society. Hydrogel-based multifunctional dressing has shown strong potential in the treatment of acute and chronic wounds. In addition to its good histocompatibility, cell adhesion, and biodegradability, methacrylic anhydride gelatin (GelMA) hydrogel has also attracted much attention due to its low cost, mild reaction conditions, adjustable physicochemical properties, and wide clinical applications. In this paper, the characteristics of GelMA hydrogel and its research progress in wound repair are introduced, and the future development of multifunctional GelMA hydrogel dressing for wound treatment is prospected.
Humans ; Gelatin/chemistry* ; Hydrogels ; Anhydrides ; Quality of Life ; Methacrylates/chemistry*

The purpose of this study was to evaluate the color stability of light cured composite resins according to the resin matrices. Three kinds of resin matrices such as Bis-GMA (Venus® Diamond; VD), Silorane (FiltekTM P90; P90), and Ormocer (Admira; Ad) were selected. Each of them were used to prepare 30 specimens. The initial CIE L*a*b* values were measured by means of a Spectrophotometer (NF999, Nippon Denshoku, Japan). For accelerated aging process, the specimens were immersed in a circulating water bath with distilled water at 60℃. Spectrophotometric analyses were made after immersion of 8, 16, 24, 41 days and the color difference (ΔE*) was calculated. The L* value increased compared with initial L* value. VD had the greatest L* value change and Ad had the smallest L* value change (p<0.05). Ad had the greatest a* value change and VD had the smallest a* value change (p<0.05). The b* value of P90 increased steadily in positive value (+)(p<0.05). ΔE* of Ad had the smallest value and that of P90 had the greatest change with time. According to this study, the value of light cured composite resins was increased with water sorption and color of those changed into greenish and yellowish. And according to the evaluation of color stability, Ad with Ormocer as resin matrices had the best color stability with time in oral environment and P90 with Silorane as resin matrices had the lowest color stability with time. And VD had the greatest initial ΔE*.
Aging ; Baths ; Bisphenol A-Glycidyl Methacrylate ; Composite Resins ; Diamond ; Immersion ; Organically Modified Ceramics ; Silorane Resins ; Water

A diblock copolymer, poly(ethylene glycol) methacrylate-block-glycidyl methacrylate (PEGMA-GMA), was prepared on glass substrate by surface-initiated atom transfer radical polymerization (SI-ATRP), and endothelial specific peptide Arg-Glu-Asp-Val (REDV) was immobilized at the end of the PEGMA-GMA polymer brush by ring opening reaction through the rich epoxy groups in the GMA. The structure and hydrophilicity of the polymer brushes were characterized by static water contact angle, X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). The results showed that the REDV modified copolymer brushes were successfully constructed on the glass substrates. The REDV peptide immobilized onto surface was quantitatively characterized by ultraviolet-visible spectroscopy (UV-VIS). The blood compatibility of the coating was characterized by recalcification time and platelet adhesion assay. The results showed that the polymer coating had good blood compatibility. The multifunctional active polymer coating with PEGMA and peptide produced an excellent prospect in surface construction with endothelial cells selectivity.
Biocompatible Materials ; Cells, Cultured ; Endothelial Cells ; Glass ; Humans ; Immobilized Proteins ; Methacrylates ; Oligopeptides ; Platelet Adhesiveness ; Polyethylene Glycols ; Polymers ; Surface Properties

Biofilms at the tooth-restoration bonded interface can produce acids and cause recurrent caries. Recurrent caries is a primary reason for restoration failures. The objectives of this study were to synthesize a novel bioactive dental bonding agent containing dimethylaminohexadecyl methacrylate (DMAHDM) and 2-methacryloyloxyethyl phosphorylcholine (MPC) to inhibit biofilm formation at the tooth-restoration margin and to investigate the effects of water-aging for 6 months on the dentin bond strength and protein-repellent and antibacterial durability. A protein-repellent agent (MPC) and antibacterial agent (DMAHDM) were added to a Scotchbond multi-purpose (SBMP) primer and adhesive. Specimens were stored in water at 37 °C for 1, 30, 90, or 180 days (d). At the end of each time period, the dentin bond strength and protein-repellent and antibacterial properties were evaluated. Protein attachment onto resin specimens was measured by the micro-bicinchoninic acid approach. A dental plaque microcosm biofilm model was used to test the biofilm response. The SBMP + MPC + DMAHDM group showed no decline in dentin bond strength after water-aging for 6 months, which was significantly higher than that of the control (P < 0.05). The SBMP + MPC + DMAHDM group had protein adhesion that was only 1/20 of that of the SBMP control (P < 0.05). Incorporation of MPC and DMAHDM into SBMP provided a synergistic effect on biofilm reduction. The antibacterial effect and resistance to protein adsorption exhibited no decrease from 1 to 180 d (P > 0.1). In conclusion, a bonding agent with MPC and DMAHDM achieved a durable dentin bond strength and long-term resistance to proteins and oral bacteria. The novel dental bonding agent is promising for applications in preventive and restorative dentistry to reduce biofilm formation at the tooth-restoration margin.
Anti-Infective Agents ; chemistry ; pharmacology ; Biofilms ; drug effects ; Dental Bonding ; Dentin-Bonding Agents ; chemistry ; pharmacology ; Materials Testing ; Methacrylates ; chemistry ; pharmacology ; Phosphorylcholine ; analogs & derivatives ; chemistry ; pharmacology ; Resin Cements ; Shear Strength ; Surface Properties ; Water

OBJECTIVE: This investigation aimed to develop a novel self-healing and antibacterial dental resin composite. The effects of water immersion on its properties were also evaluated. METHODS: Microcapsules filled with healing agent of triethylene glycol dimethacrylate were synthesized on the basis of previous studies. Antibacterial resin composite contained nano-antibacterial inorganic fillers that were modified by quaternary ammonium salt with long-chain alkyl. Microcapsules were incorporated into antibacterial resin composite at mass fraction of 7.5%. A commercial resin composite named Tetric N-Ceram was used as control. The resin samples were immersed in 37 °C distilled water for different periods. A flexural test was used to measure the mechanical properties of the novel resin composite. A single-edge V-notched beam method was used to measure fracture toughness and self-healing efficiency. A dental plaque microcosm biofilm model with human saliva as inoculum was formed. Colony-forming units (CFU) and lactic acid production of biofilm on the novel resin composite were calculated to test the antibacterial property. RESULTS: Mechanical properties and fracture toughness decreased significantly after the composite was immersed in water for 30 days (P<0.05), and no significant reduction was found from then on (P>0.05). Water immersion did not weaken the self-healing capability of the composite (P>0.05), and self-healing efficiency of 64% could still be obtained even after 270 days. The antibacterial resin composite showed a strong inhibition effect on the biofilm metabolic activity versus water immersion time from 1 day to 270 days. Therefore, the composite could still have a promising antibacterial property even after being immersed in water (P<0.05). CONCLUSIONS Water immersion could weaken the mechanical properties of the novel self-healing and antibacterial resin composite, but it insignificantly affected the self-healing and antibacterial properties of the composite.
Anti-Bacterial Agents ; Biofilms ; Capsules ; Composite Resins ; Dental Plaque ; Humans ; Materials Testing ; Methacrylates ; Water

OBJECTIVE: This investigation aimed to develop a novel antibacterial dental adhesive containing nanoantibacterial inorganic fillers and measure the dentin bonding strength, mechanical properties, and antibacterial property of the novel adhesive in vitro. METHODS: Novel nanoantibacterial inorganic fillers containing quaternary ammonium salt with long chain alkyl were synthesized on the basis of previous research. These novel nanoantibacterial inorganic fillers were added into the dental adhesive to prepare novel nanoantibacterial dental resin composite at mass fractions of 0%, 2.5%, 5.0%, 7.5%, and 10%; 0% was used as control. Dentin shear bonding test was used to evaluate the bonding strength. Flexural test was utilized to measure the novel resin composite flexural strength and elastic modulus. A dental plaque microcosm biofilm model with human saliva as inoculum was formed. Colony forming unit, lactic acid production, and live/dead assay of the biofilm on novel dental adhesive were calculated to assess the effect of novel dental adhesive on human dental plaque microcosm biofilm. RESULTS: The dentin shear bond strength, flexural strength, and elastic modulus were 28.9 MPa, 86.6 MPa, and 4.2 GPa, respectively, when the nanoantibacterial inorganic filler mass fraction in the dental adhesive reached approximately 5.0%. Consequently, the dentin shear bond strength and mechanical properties significantly increased. Addition of 2.5% nanoantibacterial inorganic fillers into the dental adhesive exerted no adverse effect on the mechanical properties significantly (P>0.05). Dental adhesive containing 5% or more nanoantibacterial inorganic fillers inhibited the metabolic activity of the dental plaque microcosm biofilm significantly, thereby displaying a strong antibacterial potency (P<0.05). CONCLUSIONS This novel antibacterial dental adhesive, which contained 5.0% nanoantibacterial inorganic filler, exhibited promising bonding strength, mechanical property, and antibacterial ability. Hence, this adhesive can be potentially used in caries inhibition in dental application.
Anti-Bacterial Agents ; Biofilms ; Dental Bonding ; Dental Cements ; Dental Plaque ; prevention & control ; Humans ; Materials Testing ; Methacrylates ; Resin Cements ; Shear Strength

Continuing advances in dentin bonding technology and adhesives revolutionized bonding of resin-based composite restorations. However, hybrid layers created by contemporary dentin adhesives present imperfect durability, and degradation of collagen matrix by endogenous enzymes is a significant factor causing destruction of hybrid layers. Bond durability can be improved by using enzyme inhibitors to prevent collagen degradation and to preserve integrity of collagen matrix. This review summarizes progress on matrix metalloproteinase inhibitors (including chlorhexidine, ethylenediaminetetraacetic acid, quaternary ammonium salt, tetracycline and its derivatives, hydroxamic acid inhibitors, bisphosphonate derivative, and cross-linking agents) and suggests prospects for these compounds.
Acid Etching, Dental ; Bisphenol A-Glycidyl Methacrylate ; Collagen ; Dental Bonding ; Dentin ; Dentin-Bonding Agents ; Humans ; Matrix Metalloproteinase 2 ; Matrix Metalloproteinase Inhibitors