Experimental Study on Human Demineralized Dentin Matrix as rhBMP-2 Carrier <em>In Vivo</em>

Research Article

J Dent App. 2015; 2(7): 269-273.

Experimental Study on Human Demineralized Dentin Matrix as rhBMP-2 Carrier In Vivo

In-Woong Um¹*, Woo-Jin Cho² and Young-Kyun Kim³

¹Director, R&D Institute, Korea Tooth Bank, Seoul, Korea

²Researcher, R&D Institute, Korea Tooth Bank, Seoul, Korea

³Professor, Department of Oral and Maxillofacial Surgery, Section of Dentistry, Seoul National University Bundang Hospital, Seongnam, Korea

*Corresponding author: In-Woong Um, DDS, PhD, CTBS, R&D Institute Korea Tooth Bank, Seoul-In Dental Clinic, Korea

Received: June 16, 2015; Accepted: September 10, 2015; Published: September 12, 2015


Purpose: This study examined the bone induction capacity of human demineralized dentin matrix as an rhBMP-2 carrier.

Method: Total 40 nude mice weighing 15-20g were used for the study. Two types of scaffold were selected as a carrier of rhBMP-2: Inorganic bovine bones, tricalcium phosphate (TCP) as the control group where Autogenous tooth bone grafts material (AutoBT), a human demineralized dentin matrix (DDM), as the experimental group. Laboratory animals were sacrificed at two and four weeks after the implanting two rhBMP-2 carriers into intramuscular pouch. The evaluation of the specimen was divided into four compartments: field phenomenon, cell number (osteoblast, firbroblast), osteoid formation, and maturation, and compared two carriers histologically.

Result: Field phenomenon: Both groups have excellent fibrous encapsulations around particles; Cell number (osteoblast, fibroblast): The early cellular reaction on the surface of particles was superior in the DDM group; Osteoid formation: more osteoid was deposited on the DDM group; Maturation: The DDM group showed compact osteoid bridges and dense fibrous connective tissue filled between each particle, but the TCP group showed that the ostoid deposited seemed to be transformed into fatty tissues after 4 weeks.

Conclusion: Both the DDM/rhBMP-2 and TCP/rhBMP-2 induced bone in intramuscular pouch of nude mice, but more bone was formed in DDM/rhBMP-2 group.

Keywords: Bone induction; Demineralized dentin matrix; DDM; AutoBT


DDM: Demineralized Dentin Matrix; AutoBT: Autogenous Tooth Bone Graft Material; Inorganic Bovine Bones; TCP: Tricalcium Phosphate; BMP: Bone Morphogenetic Protein.


In 1967, there was a report that rabbit demineralized matrices (DDM) induced bone formation in the intramuscular pockets [1,2]. Since the result indicated that Bone Morphogenetic Proteins (BMPs) were involved in dentin matrix, there have been several studies to examine the animal DDM could induce ectopic bone formation in subcutaneous and intramuscular pockets in rodents [3,4].

In 2005, human DDM particles finally succeeded to induce ectopic bone and cartilage [5]. And continuous researches had developed human autogenous demineralized dentin matrix (AutoBT) that has been actively used in Korea and Japan for the past 5 years [6]. Also, another report indicated that DDM induced bone and cartilage independently while demineralized bone matrix (DBM) induced cartilage, bone and marrow at 4 weeks in the back skin of nude mice [7]. Lastly, there was a report that bone and cartilage was induced in nude mice using AutoBT by itself [8].

On the other side, a report in 1998 suggested that root dentin prepared from extracted teeth could be recycled for using as a carrier of rhBMP-2 because it induces new bone formation in the periodontium while another report showed that the osteoinductive matrices of human DDM particles could be effective as a carrier of rhBMP-2 for bone engineering later in 2005 [5, 9].

Therefore, the purpose of this study is to evaluate the bone induction capacity of DDM when fixed with rhBMP-2 whether DDM could act as anrhBMP-2 carrier.

Materials and Methods

Two types of scaffold were selected as a carrier of rhBMP-2 (CowellMedi): AutoBT (human DDM, Korea Tooth Bank, Seoul, Korea) and TCP (CowellMedi, Busan, Korea). rhBMP-2-soaked TCP (TCP/rhBMP-2) was used for the control group (n=20) where rhBMP-2-soaked DDM (DDM/rhBMP-2) was the experimental group (n=20).

Preparations of DDM

An extracted human tooth was soaked in 70% ethyl alcohol and cleaned by removing foreign substances such as restoration materials, root canal and prosthetics as well as soft tissues of periodontium, pulp and caries. After dividing cleaned tooth into crown and root, the root portion was collected and prepared for partially demineralized dentin matrix (DDM) as reported previously.0.3 to 0.8 mm crushed particles were soaked in distilled water and hydrogen dioxide solution and the remaining foreign substances were removed by ultrasonic cleaner. The cleaned particles were dehydrated with ethyl alcohol and went through defatting using ethyl ether solution. The particles were then decalcified for 2 hours in 0.6 N HCl by repeated30-minute periods [8,10]. After the demineralization, the dentinal tubules and compact peritubular dentinal collagens are widened and loosened respectively (Figure 1A, 1B).