The present invention relates to preparation of bioink composed of cellulose nanofibril hydrogel with native or synthetic Calcium containing particles. The concentration of the calcium containing particles can be between 1% and 40% w/v. Such bioink can be 3D Bioprinted with or without human or animal cells. Coaxial needle can be used where cellulose nanofibril hydrogel filled with Calcium particles can be used as shell and another hydrogel based bioink mixed with cells can be used as core or opposite. Such 3D Bioprinted constructs exhibit high porosity due to shear thinning properties of cellulose nanofibrils which provides excellent printing fidelity. They also have excellent mechanical properties and are easily handled as large constructs for patient-specific bone cavities which need to be repaired. The porosity promotes vascularization which is crucial for oxygen and nutrient supply. The porosity also makes it possible for further recruitment of cells which accelerate bone healing process. Calcium containing particles can be isolated from autologous bone, allogenic bone or xenogeneic bone but can be also isolated from minerals or be prepared by synthesis. Preferable Calcium containing particles consist of β-tricalcium phosphate which is resorbable or natural bone powder, preferably of human or porcine origin. The particles described in the present invention have particle size smaller than 400 microns, or more preferably smaller than 200 microns, to make it possible to handle in printing nozzle without clogging and to obtain a good resolution. Cellulose nanofibrils can be produced by bacteria orbe isolated from plants. They can be neutral, charged or oxidized to be biodegradable. The bioink can be additionally supplemented by other biopolymers which provide crosslinking. Such biopolymers can be alginates, chitosans, modified hyaluronic acid or modified collagen derived biopolymers.

A dentin regenerative cell culture that can bring about a rapid regeneration of dentin in a deficit region. In the treatment of a tooth using the dentin regenerative cell culture, a root canal where a pulpectomy has been performed is filled with a root canal filler containing dental pulp stem cells. The dentin regenerative cell culture is then implanted in the deficit region of dentin, and temporary sealing with a packing is carried out. The dentin regenerative cell culture is formed three-dimensionally in conformity with the shape of the deficit region with the coalescence of cell masses of a plurality of odontoblasts, and thus the dentin regeneration is well promoted. In addition, gaps between the dentin regenerative cell culture and biological tissue can be rapidly filled. Infection due to bacterial infiltration can thereby be prevented.

Hydrogel-based scaffolds useful for promoting pulp cell growth and biosynthesis, regulating pulp cell migration and morphology, or both as well as methods for their production and use are provided.

Provided is a method for making a biotransplant, comprising introducing autologous fibroblasts isolated from an oral mucosa of a patient into a platelet-rich fibrin (PRF) membrane using linear retrograde needle injection, where a needle is inserted into a thickness of the PRF membrane and a first puncture is made and then a series of punctures are made that are linearly aligned or arrayed with the first puncture and are spaced at a predetermined distance from a prior puncture. A method of treatment of a periodontal tissue and a biotransplant comprising a PRF membrane and autologous fibroblasts are also provided.

A dental hydrogel composition comprising: (a) polysaccharide; (b) polydopamine conjugated to the polysaccharide, wherein between 5 and 35 percent of polysaccharide sugar moieties are conjugated to polydopamine; (c) RGD peptide coupled to the polysaccharide-poly-dopamine conjugate; and (d) moieties that are crosslinkable upon exposure to light coupled to the polysaccharide; wherein components (a)-(d) are disposed in the composition such that the hydrogel composition: exhibits an adhesive strength of at least 10 kPa upon cross linking of crosslinkable moieties; and exhibits an elasticity between 5 kPa and 100 kPa upon cross linking of crosslinkable moieties.

The object of the present disclosure is to achieve a composite of dentin collagen obtained from human teeth and a growth factor with function of inducing bone growth, thereby improving a regeneration speed of alveolar bone. The present disclosure provides a method for manufacturing a bone graft composite, said method comprising: pulverizing a tooth to generate tooth powder, and washing and degreasing the tooth powder; demineralizing the degreased tooth powder so as to obtain dentin collagen; preparing a growth factor which includes a component to induce bone growth; and synthesizing a composite by mixing the obtained dentin collagen and growth factor.

[Problem] The present invention addresses the problem of providing a method for efficiently preparing dental pulp stem cells from dental pulp cells collected from dental pulp tissue.

[Solution Means] The method for preparing dental pulp stem cells from dental pulp cells collected from dental pulp tissue according to the present invention includes: (a) a step for preculturing dental pulp cells using a culture medium that includes serum at 7-13% (v/v), the dental pulp cells being bonded to a culture dish; (b) a step for culturing the dental pulp cells bonded to the culture dish by step (a) at least for one day using a culture medium that includes serum at 7-13% (v/v); and (c) a step for culturing the dental pulp cells in a low-serum condition after step (b).

A non-cellular dental tissue regeneration promoting kit is provided that makes effective dental tissue regeneration possible without transplantation of autologous or allogeneic stem cells or components derived from stem cells. The kit comprises: a pretreatment agent comprising a serine protease; and a non-cellular root canal filling material, the non-cellular root canal filling material comprising: a regeneration promoting compound including at least one of a CCR3 antagonist, a CCL11 neutralizing antibody, and an ALK5 inhibitor; and an extracellular matrix. The kit is preferably used for middle-aged or elderly individuals.

Provided is a dental pretreatment material for dental tissue regeneration by use of dental pulp stem cells, particularly a dental pretreatment material effectively enabling dental tissue regeneration even by use of dental pulp stem cells of middle-aged or older individuals. The dental pretreatment material is characterized by comprising a serine protease, specifically trypsin. The dental pretreatment material comprising trypsin is used as an injection into a root canal before a root canal filling material comprising dental pulp stem cells and an extracellular matrix is inserted into the root canal as an attempt to regenerate a dental pulp and a dentin. The root canal filling material includes an ALK5 inhibitor, a CCR3 antagonist, or a CCL11 neutralizing antibody.

The present invention provides a dental implant device comprising a dental implant body and an implant fixture. Said implant fixture is characterized in that it comprises a portion to be fixed to said implant body and a portion to be fixed to a tooth adjacent to the transplantation site of said implant body in a subject, and is configured so that mechanical stimulation from the tooth adjacent to said transplantation site is transmissible to said implant body, and said implant device is characterized in that it is placed into the oral cavity of said subject so that said implant body is inserted at said transplantation site while said implant fixture is fixed to said implant body and the tooth adjacent to said transplantation site.