Effects of the incorporation of ε-aminocaproic acid/chitosan particles to fibrin on cementoblast differentiation and cementum regeneration

Park, C.H.,Oh, J.H.,Jung, H.M.,Choi, Y.,Rahman, S.U.,Kim, S.,Kim, T.I.,Shin, H.I.,Lee, Y.S.,Yu, F.H.,Baek, J.H.,Ryoo, H.M.,Woo, K.M. Elsevier Science B.V. Amsterdam 2017 ACTA BIOMATERIALIA Vol. No.

Cementum formation on the exposed tooth-root surface is a critical process in periodontal regeneration. Although various therapeutic approaches have been developed, regeneration of integrated and functional periodontal complexes is still wanting. Here, we found that the OCCM30 cementoblasts cultured on fibrin matrix express substantial levels of matrix proteinases, leading to the degradation of fibrin and the apoptosis of OCCM30 cells, which was reversed upon treatment with a proteinase inhibitor, ε-aminocaproic acid (ACA). Based on these findings, ACA-releasing chitosan particles (ACP) were fabricated and ACP-incorporated fibrin (fibrin-ACP) promoted the differentiation of cementoblasts in vitro, as confirmed by bio-mineralization and expressions of molecules associated with mineralization. In a periodontal defect model of beagle dogs, fibrin-ACP resulted in substantial cementum formation on the exposed root dentin in vivo, compared to fibrin-only and enamel matrix derivative (EMD) which is used clinically for periodontal regeneration. Remarkably, the fibrin-ACP developed structural integrations of the cementum-periodontal ligament-bone complex by the Sharpey's fiber insertion. In addition, fibrin-ACP promoted alveolar bone regeneration through increased bone volume of tooth roof-of-furcation defects and root coverage. Therefore, fibrin-ACP can promote cementogenesis and osteogenesis by controlling biodegradability of fibrin, implicating the feasibility of its therapeutic use to improve periodontal regeneration. Statement of Significance: Cementum, the mineralized layer on root dentin surfaces, functions to anchor fibrous connective tissues on tooth-root surfaces with the collagenous Sharpey's fibers integration, of which are essential for periodontal functioning restoration in the complex. Through the cementum-responsible fiber insertions on tooth-root surfaces, PDLs transmit various mechanical responses to periodontal complexes against masticatory/occlusal stimulations to support teeth. In this study, periodontal tissue regeneration was enhanced by use of modified fibrin biomaterial which significantly promoted cementogenesis within the periodontal complex with structural integration by collagenous Sharpey's fiber insertions in vivo by controlling fibrin degradation and consequent cementoblast apoptosis. Furthermore, the modified fibrin could improve repair and regeneration of tooth roof-of-furcation defects, which has spatial curvatures and geometrical difficulties and hardly regenerates periodontal tissues.

Hyaluronic Acid가 첨가된 Fibrin의 연골 세포 전달 지지체 제작

민병현,박상혁,박소라,박근홍,정수일 한국생체재료학회 2004 생체재료학회지 Vol.8 No.1

Tissue engineering is the process of creating functional biologic prostheses by suspending dissociated cells in biodegradable polymer scaffolds, Various attempts have been described that demonstrate the suitability of cultured chondrocytes in gel substances like agarose, hyalronate (HA), fibrin glue, collagen, alginate. However, these gel cultures do not provide suitable mechanical stability necessary for cartilage constructs with a specific predefined shape for transplantation. In this study fibrin/HA composite gel has been used for cell delivery vehicle which showed the excellent scaffold because fibrin can occur a substantial reduction of mass after in vivo implantation. Such an adhesive composite gel could provide ready made cell delivery vehicle for transplant fixation and may allow a permissive microenvironment. In stable fibrin gel intermeshed with high levels of HA, Fibrin/HA composite gel was able to support growth and differentiation of a high number of chondrocytes in vitro. The cell density (5?06 cells/ml) was selected in fibrin/HA composite gel because of it appropriate cell number for chondrogenesis. In addition, the fibrinolysis inhibition factor (FIF) was mixed with fibrin/HA composite gel in order to inhibit gel degradation. The addition of FIF led to larger cartilage formation. Therefore, we retested superior of fibrin/HA gel construct. Cartilage formation was larger in the fibrin/HA+FIF gel than in fibrin/HA gel. These results supported the feasibility of tissue-engineered cartilage using both fibrin and HA as candidates of extracelular matrix.

Tissue-engineered Cartilage Using Fibrin/Hyaluronan Composite Gel and Its In Vivo Implantation

Park, Sang-Hyug,Park, So Ra,Chung, Soo Il,Pai, Ki Soo,Min, Byoung-Hyun Blackwell Science Inc 2005 Artificial Organs Vol.29 No.10

<P>Abstract: </P><P>The importance of scaffold biomaterials has been emphasized for in vitro culture of tissue-engineered cartilage in a three-dimensional (3D) environment. In this study, we examined the feasibility of fibrin glue, mixed with hyaluronic acid (HA) as a composite scaffold. Fibrin glue has been a useful cell delivery matrix for cartilage tissue engineering and HA is a key component of normal articular cartilage. Our hypothesis is that compared to fibrin itself, a fibrin/HA composite can have significantly enhanced properties, due mainly to the added benefits of HA in the matrix. Pieces of cartilage were isolated from rabbit knees and the chondrocytes were harvested through enzymatic digestion. Both fibrin and fibrin/HA composite were prepared and subsequently implanted in nude mice (<I>n</I> = 9, each group) for 1, 2, and 4 weeks, respectively. The retrieved specimens were then analyzed and the results were compared. Cartilage-like tissue formation was detected earlier with fibrin/HA specimens. They produced significantly higher amounts of the extracellular matrix (ECM) molecules, GAG, and collagen at each time point than those in fibrin. Interestingly, the fibrin/HA composite was also competent in maintaining its initial size. Histology—Safranin O/fast green and Alcian blue—of the retrieved specimens found more intense, uniform staining in the fibrin/HA composites. Analysis of the gene expression of the ECM molecules also confirmed the benefits of the composite with added HA in the maintenance of phenotypic stability. The present study suggests that fibrin/HA composite may serve as a dependable cell delivery vehicle as well as a structural basis for tissue-engineered cartilage.</P>

Fibrin-Enhanced Canonical Wnt Signaling Directs Plasminogen Expression in Cementoblasts

Rahman, Saeed Ur,Park, Chan Ho,Baek, Jeong-Hwa,Ryoo, Hyun-Mo,Woo, Kyung Mi MDPI AG 2017 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.18 No.11

<P>Cementum is a mineralized layer on the tooth’s root surface and facilitates the biomechanical anchoring of fibrous connective tissues as a part of tooth-supportive complexes. Previously, we observed that OCCM30 cementoblasts cultured on fibrin matrices underwent apoptosis due to fibrin degradation through the expression of proteases. Here, we demonstrated that OCCM30 on fibrin matrices (OCCM30-fibrin) enhanced canonical Wnt signaling, which directed to plasminogen expression. The OCCM30-fibrin showed higher levels of Wnt3a expression, nuclear translocation of β-catenin, and T-cell factor (TCF) optimal motif (TOP) reporter activity than the cells on tissue culture dishes (OCCM30-TCD), indicating that the OCCM30-fibrin enhanced canonical Wnt/β-catenin signaling. Also, OCCM30-fibrin expressed biomineralization-associated markers at higher levels than OCCM30-TCD, of which levels were further increased with LiCl, a Wnt signaling activator. The OCCM30 cementoblasts simultaneously showed that high levels of plasminogen, a critical component of fibrinolysis, were expressed in the OCCM30-fibrin. Activation of canonical Wnt signaling with LiCl treatment or with forced lymphoid enhancer factor 1 (LEF1)-expression increased the expression of plasminogen. On the contrary, the inhibition of canonical Wnt signaling with siRNAs against Wnt3a or β-catenin abrogated fibrin-enhanced plasminogen expression. Furthermore, there are three conserved putative response elements for the LEF1/β-catenin complex in the plasminogen proximal promoter regions (−900 to +54). Site-directed mutations and chromatin immunoprecipitation indicated that canonical Wnt signaling directed plasminogen expression. Taken together, this study suggests that fibrin-based materials can modulate functional periodontal formations in controlling cementoblast differentiation and fibrin degradation.</P>

Biomimetic chimeric peptide-tethered hydrogels for human mesenchymal stem cell delivery

Shim, G.,Kim, G.,Choi, J.,Yi, T.,Cho, Y.K.,Song, S.U.,Byun, Y.,Oh, Y.K. Elsevier 2015 Colloids and Surfaces B Vol.136 No.-

Here, we report a chimeric peptide-tethered fibrin hydrogel scaffold for delivery of human mesenchymal stem cells (hMSC). Osteopontin-derived peptide (OP) was used as an hMSC-tethering moiety. OP showed hMSC adhesion properties and enhanced hMSC proliferation. A natural fibrin-binding protein-derived peptide (FBP) was tested for its ability to tether hMSC to the fibrin gel matrix. FBP loading on fibrin gels was 8.2-fold higher than that of a scrambled peptide (scFBP). FBP-loaded fibrin gels were retained at injection sites longer than scFBP-loaded fibrin gels, showing a 15.9-fold higher photon intensity of fluorescent FBP-grafted fibrin gels than fluorescent scFBP-loaded fibrin gels 48h after injection. On the basis of the fibrin gel-binding properties of FBP and the hMSC-binding and proliferation-supporting properties of OP, we constructed chimeric peptides containing FBP and OP linked with a spacer (FBPsOP). Four days after transplantation, the survival of hMSC in FBPsOP-grafted fibrin gels was 3.9-fold higher than hMSC in fibrin gels alone. Our results suggest the potential of FBPsOP-grafted fibrin gels as a bioactive delivery system for enhanced survival of stem cells.

중이수술에 인체에서 추출한 Fibrin 접착제의 이용 : Ammonium Sulfate fibrin 접착제와 ??의 비교

이형철,양미경,박문흠 영남대학교 의과대학 1991 Yeungnam University Journal of Medicine Vol.8 No.1

성공적인 종이수술을 위하여 안전하고 효과적인 접착제가 필요하며, 합성 접착제의 독성작용 때문에 인체에서 추출한 접착제를 사용하게 되었다. Fibrin 접착제가 저장 혈액에서 체취되면 감염성 질환의 전염 위험성이 있지만 자가 Fibrin접착제를 사용하면 전염위험성이 없다. 저자들은 ammonium sulfate fibrin 접착제와 상품화된 fibrin 접착제의 접착력을 비교한 결과 ammonium sulfate fibrin 접착제의 접착력이 상품화된 fibrin 접착제의 반 정도의 접착력을 가지지만 그 정도의 접착력이면 고막재생술, 안면신경 봉합술, 이소골 재건술, 외이도 후벽재건술, 뼈가루를 이용한 전두동 및 유양동폐쇄술등 이비인후과 수술에 사용하기엔 충분한 접착력이다. Successful middle ear surgery requires the availability of al safe, effective bonding material. Side effect caused by synthetic materials have led to the use of biologic adhesive, However, they carry the risk of transmission of infectious diseases if they are prepared from pooled human blood. The adhesive strength of ammonium sulfate fibrin adhesive produce an adhesive strength that is half that of the homologous commercial product. It is, however, good enough for use in several otolaryngological operations, tympanoplasty, facial nerve repair, reconstruction of ossicles. reconstruction of posterior wall of ear canal and obliteration of frontal sinus and mastoid antrum using bone dust.

감염된 미성숙 영구치에서 platelet-rich fibrin과 double antibiotic paste를 이용한 치수 재혈관화 : 증례 보고

전상윤,이난영,이상호 大韓小兒齒科學會 2013 大韓小兒齒科學會誌 Vol.40 No.3

Paradigm shift in management of infected immature permanent teeth has occurred. The new concept of the treatment includes minimal or no intracanal instrumentation, disinfection with triple antibiotic paste and sealing with mineral trioxide aggregate. This regenerative endodontic treatment promotes differentiation of periradicular stem cells that induce regeneration of vital tissue and continuation of root formation. Thorough disinfection and three-dimensional scaffold are important in this new concept of the treatment. Platelet-rich fibrin has been reported as 'new scaffold' instead of blood clot, which had been used in the past. Triple antibiotics can be used to disinfect the tooth but may lead to complications including discoloration. Three cases of infected immature permanent tooth caused by dens evaginatus fracture are presented. After removal of necrotic pulp and thorough intracanal irrigation, only platelet-rich fibrin was applied to the root canal in the first case. In the other cases, topical antibiotics was used for disinfection and platelet-rich fibrin for scaffold. In all the cases, the opening was sealed with mineral trioxide aggregate. All the cases showed proper healing of inrabony lesion and some lengthening of root. According to these cases, regenerating vital tissue of the infected immature permanent tooth can be achieved with disinfection and application of platelet-rich fibrin. 감염된 미성숙 영구치의 치수치료에 있어 줄기세포의 분화를 유도하는 생활조직의 재생과 지속적인 치근형성을 도모하는방향으로 패러다임이 전환되고 있는데, 여기에서는 소독, 스캐폴드(scaffold), 그리고 폐쇄가 중요하다. 소독을 위해 triple antibiotics가 널리 사용되고 있으며, 스캐폴드로써 기존의 혈병대신 platelet-rich fibrin의 사용이 보고되었다. 본 증례보고에서는 치외치 파절에 의해 치수가 감염된 미성숙 영구치에서 platelet-rich fibrin을 스캐폴드로써 이용한 치수 재혈관화를시행하였다. 발수와 근관세척 후 첫 증례에서 국소적 항생제의 적용 없이 platelet-rich fibrin을 단독 사용하였고 두 번째와세 번째 증례에서는 국소적 항생제 적용 후 platelet-rich fibrin을 적용하였는데 모두 양호한 치유 결과를 얻었다.

Potential of Fortified Fibrin/Hyaluronic Acid Composite Gel as a Cell Delivery Vehicle for Chondrocytes

Park, Sang-Hyug,Cui, Ji Hao,Park, So Ra,Min, Byoung-Hyun Blackwell Publishing Inc 2009 Artificial Organs Vol.33 No.6

<P>Abstract</P><P>Numerous treatment methods have been applied for use in cartilage repair, including abrasion, drilling, and microfracture. Although chondrocyte transplantation is the preferred treatment, it has some shortcomings, such as difficulty of application (large and posterior condylar regions), poor chondrocyte distribution, and potential cell leakage from the defect region. The cell delivery system of the tissue engineering technique can be used to overcome these shortcomings. We chose fibrin/hyaluronan (HA) composite gel as an effective cell delivery system to resolve these issues. Both components are derived from natural extracellular matrix. In the first trial, fortified fibrin/HA composite gels with rabbit chondrocytes were tested by implantation in nude mice. At 4 weeks, glycosaminoglycan contents in the fibrin/HA composite (0.186 ± 0.006 mg/mg) were significantly higher than those in the presence of fibrin alone (0.153 ± 0.017 mg/mg). As a next step, we applied the fibrin/HA composite gel to animal cartilage defects using full thickness cartilage defect rabbit models. The fibrin/HA composite gel with rabbit chondrocytes (allogenic) was implanted into the experimental group, and the control group was implanted with the fibrin/HA composite gel alone. Implanted chondrocytes with the fibrin/HA composite showed improved cartilage formation. In conclusion, the key to successful regeneration of cartilage is to provide the repair site with a sufficient supply of chondrogenic cells with a suitable delivery vehicle to ensure maximal differentiation and deposition of the proper extracellular matrix. This study suggests the feasibility of tissue-engineered cartilage formation using fibrin/HA composite gel.</P>

Fibrin affects short-term in vitro human mesenchymal stromal cell responses to magneto-active fibre networks

Spear, Rose L.,Symeonidou, Antonia,Skepper, Jeremy N.,Brooks, Roger A.,Markaki, Athina E. Techno-Press 2015 Biomaterials and biomedical engineering Vol.2 No.3

Successful integration of cementless femoral stems using porous surfaces relies on effective periimplant bone healing to secure the bone-implant interface. The initial stages of the healing process involve protein adsorption, fibrin clot formation and cell osteoconduction onto the implant surface. Modelling this process in vitro, the current work considered the effect of fibrin deposition on the responses of human mesenchymal stromal cells cultured on ferritic fibre networks intended for magneto-mechanical actuation of in-growing bone tissue. The underlying hypothesis for the study was that fibrin deposition would support early stromal cell attachment and physiological functions within the optimal regions for strain transmission to the cells in the fibre networks. Highly porous fibre networks composed of 444 ferritic stainless steel were selected due to their ability to support human osteoblasts and mesenchymal stromal cells without inducing untoward inflammatory responses in vitro. Cell attachment, proliferation, metabolic activity, differentiation and penetration into the ferritic fibre networks were examined for one week. For all fibrin-containing samples, cells were observed on and between the metal fibres, supported by the deposited fibrin, while cells on fibrin-free fibre networks (control surface) attached only onto fibre surfaces and junctions. Initial cell attachment, measured by analysis of deoxyribonucleic acid, increased significantly with increasing fibrinogen concentration within the physiological range. Despite higher cell numbers on fibrin-containing samples, similar metabolic activities to control surfaces were observed, which significantly increased for all samples over the duration of the study. It is concluded that fibrin deposition can support the early attachment of viable mesenchymal stromal cells within the inter-fibre spaces of fibre networks intended for magneto-mechanical strain transduction to in-growing cells.

다공성 안와충전물에서 혈관생성 및 유지에 대한 혈관전구세포와 섬유소의 역할

양재욱,이호영,박세광,양영일.Jae Wook Yang. M.D.. Ph.D.. Ho Young Lee. M.D.. Sae Gwang Park. M.D.. Ph.D.. Young Il Yang. M.D.. Ph.D. 대한안과학회 2008 대한안과학회지 Vol.49 No.7

Purpose: The effects of endothelial progenitor cells (EPCs) and fibrin on fibrovascular growth into porous polyethylene orbital implants (Medpor® sheet) were investigated using stem cells. Methods: EPCs were separated from human adipose fat tissue for culture. Fluorescence-activated cell sorting (FACS) was used to identify the phenotype and to analyze the purity of EPCs cultivated from human adipose tissue. Processed Medpor® sheets were inserted in each quadrant of the subcutaneous fat layer under the dorsal surface of 20 anesthetized athymic nude mice, using sterile methods. Medpor® sheets processed with endothelial progenitor cells and fibrin were inserted into the two top quadrants, a Medpor® sheet processed with fibrin was inserted in the lower right quadrant, and an unprocessed Medpor® sheet was inserted in the lower left quadrant of each mouse. The mice were sacrificed on the seventh day. The adhesiveness and blood vessel formation were quantified by weight and the number of blood cells within the Medpor® sheets. Hematoxylin and eosin (H&E) and toluidine blue stains were used to analyze fibrovascular and cell growth within the Medpor® sheets. Results: The sheets processed with EPCs and fibrin were heavier and contained more white and red blood cells (p<0.001) than the other sheets. The sheets processed with fibrin alone were heavier (p<0.01) and contained more blood cells (p<0.001) than the unprocessed sheets. The degree of vessel formation and tissue adhesiveness was greatest in the group of Medpor® sheets processed with EPCs and fibrin. The sheets processed with fibrin only had greater tissue adhesiveness and fibrovascular proliferation than the unprocessed Medpor® sheets. Conclusions: Endothelial progenitor cells and fibrin applied to Medpor® sheets improve fibrovascular proliferation and tissue adhesiveness. When both are applied together, a synergistic effect is seen. J Korean Ophthalmol Soc 49(7):1135-1145, 2008