ROLE OF COMBINATION CASEIN AND LACTOFERRIN BOVINE’S COLLOSTRUM AS A PULP CAPPING ON MACROPHAGE EXPRESSION IN MALE WISTAR RATS

ABSTRACTBackground: Inflammatory and/or non-inflammatory processes play a role in stimulating pulp repair and the formation of hard tissue, namely reparative dentin. Macrophages play a role in the pathogenesis and chronic inflammatory disorders. The combination casein lactoferrin of bovine colostrum as an immunomodulator has therapeutic potential. This study aims to determine the therapeutic effect and duration of application of the combination of casein lactoferrin of bovine colostrum, on the expression of macrophages as pulp capping.Method: This study was a true experimental laboratories post test only control group design, consisting of three groups of 60 male wistar rats with 4 observation times, namely day to day 7, 14, 21 and 28 each of 5 mice. The maxillary 1st molars were prepared until the roof of the pulp was exposed. Three groups, namely the combination of casein and lactoferrin bovine colostrum (CKL) and calcium hydroxide (K1) and the untreated group (K0). Each group was filled with glassionomer as a permanent restoration. The tissue was made histological preparations with hematoxylin-eosin staining and the number of macrophages were counted, then analyzed by two way ANOVA and post hoc LSD tests.Result: The results showed that the therapeutic effect and duration of application of the combination of casein and lactoferrin bovine colostrum on the expression of macrophages as pulp cappingConclusion: The combination of casein and lactoferrin of bovine colostrum as capping material can increase the number of macrophages in the healing process of dental pulp.

Facilitating Reparative Dentin Formation Using Apigenin Local Delivery in the Exposed Pulp Cavity

Apigenin, a natural product belonging to the flavone class, affects various cell physiologies, such as cell signaling, inflammation, proliferation, migration, and protease production. In this study, apigenin was applied to mouse molar pulp after mechanically pulpal exposure to examine the detailed function of apigenin in regulating pulpal inflammation and tertiary dentin formation. In vitro cell cultivation using human dental pulp stem cells (hDPSCs) and in vivo mice model experiments were employed to examine the effect of apigenin in the pulp and dentin regeneration. In vitro cultivation of hDPSCs with apigenin treatment upregulated bone morphogenetic protein (BMP)- and osteogenesis-related signaling molecules such as BMP2, BMP4, BMP7, bone sialoprotein (BSP), runt-related transcription factor 2 (RUNX2), and osteocalcin (OCN) after 14 days. After apigenin local delivery in the mice pulpal cavity, histology and cellular physiology, such as the modulation of inflammation and differentiation, were examined using histology and immunostainings. Apigenin-treated specimens showed period-altered immunolocalization patterns of tumor necrosis factor (TNF)-α, myeloperoxidase (MPO), NESTIN, and transforming growth factor (TGF)-β1 at 3 and 5 days. Moreover, the apigenin-treated group showed a facilitated dentin-bridge formation with few irregular tubules after 42 days from pulpal cavity preparation. Micro-CT images confirmed obvious dentin-bridge structures in the apigenin-treated specimens compared with the control. Apigenin facilitated the reparative dentin formation through the modulation of inflammation and the activation of signaling regulations. Therefore, apigenin would be a potential therapeutic agent for regenerating dentin in exposed pulp caused by dental caries and traumatic injury.

Effectiveness of Direct Pulp Capping Bioactive Materials in Dentin Regeneration: A Review

Background: Regenerative endodontics aims to restore normal pulp function in necrotic and infected teeth, restoring protective functions, such as innate pulp immunity, pulp repair through mineralization, and pulp sensibility. The aim of this systematic review was to assess the dentin regeneration efficacy of direct pulp capping (DPC) biomaterials. Methods: The literature published between 2005 and 2021 was searched by using PubMed, Web of Science, Science Direct, Google Scholar, and Scopus databases. Clinical controlled trials, randomized controlled trials, and animal studies investigating DPC outcomes or comparing different capping materials after pulp exposure were included in this systematic review. Three independent authors performed the searches, and information was extracted by using a structured data format. Results: A total of forty studies (21 from humans and 19 from animals) were included in this systemic review. Histological examinations showed complete/partial/incomplete dentin bridge/reparative dentin formation during the pulp healing process at different follow-up periods, using different capping materials. Conclusions: Mineral trioxide aggregate (MTA) and Biodentine can induce dentin regeneration when applied over exposed pulp. This systematic review can conclude that MTA and its variants have better efficacy in the DPC procedure for dentin regeneration.

Mouse Dspp frameshift model of human dentinogenesis imperfecta

Non-syndromic inherited defects of tooth dentin are caused by two classes of dominant negative/gain-of-function mutations in dentin sialophosphoprotein (DSPP): 5′ mutations affecting an N-terminal targeting sequence and 3′ mutations that shift translation into the − 1 reading frame. DSPP defects cause an overlapping spectrum of phenotypes classified as dentin dysplasia type II and dentinogenesis imperfecta types II and III. Using CRISPR/Cas9, we generated a Dspp−1fs mouse model by introducing a FLAG-tag followed by a single nucleotide deletion that translated 493 extraneous amino acids before termination. Developing incisors and/or molars from this mouse and a DsppP19L mouse were characterized by morphological assessment, bSEM, nanohardness testing, histological analysis, in situ hybridization and immunohistochemistry. DsppP19L dentin contained dentinal tubules but grew slowly and was softer and less mineralized than the wild-type. DsppP19L incisor enamel was softer than normal, while molar enamel showed reduced rod/interrod definition. Dspp−1fs dentin formation was analogous to reparative dentin: it lacked dentinal tubules, contained cellular debris, and was significantly softer and thinner than Dspp+/+ and DsppP19L dentin. The Dspp−1fs incisor enamel appeared normal and was comparable to the wild-type in hardness. We conclude that 5′ and 3′ Dspp mutations cause dental malformations through different pathological mechanisms and can be regarded as distinct disorders.

Secreted Frizzled-Related Protein 1 Promotes Odontoblastic Differentiation and Reparative Dentin Formation in Dental Pulp Cells

Direct pulp capping is an effective treatment for preserving dental pulp against carious or traumatic pulp exposure via the formation of protective reparative dentin by odontoblast-like cells. Reparative dentin formation can be stimulated by several signaling molecules; therefore, we investigated the effects of secreted frizzled-related protein (SFRP) 1 that was reported to be strongly expressed in odontoblasts of newborn molar tooth germs on odontoblastic differentiation and reparative dentin formation. In developing rat incisors, cells in the dental pulp, cervical loop, and inner enamel epithelium, as well as ameloblasts and preodontoblasts, weakly expressed Sfrp1; however, Sfrp1 was strongly expressed in mature odontoblasts. Human dental pulp cells (hDPCs) showed stronger expression of SFRP1 compared with periodontal ligament cells and gingival cells. SFRP1 knockdown in hDPCs abolished calcium chloride-induced mineralized nodule formation and odontoblast-related gene expression and decreased BMP-2 gene expression. Conversely, SFRP1 stimulation enhanced nodule formation and expression of BMP-2. Direct pulp capping treatment with SFRP1 induced the formation of a considerable amount of reparative dentin that has a structure similar to primary dentin. Our results indicate that SFRP1 is crucial for dentinogenesis and is important in promoting reparative dentin formation in response to injury.

Endodontic Treatment of a Traumatic Tusk Fracture With Exposed Pulp in an Asian Elephant (Elephas maximus)

Tusk fracture in elephants is a common incident often resulting in pulp exposure and pulpitis. Extensive lavage, endodontic therapy, direct pulp capping, or extraction are treatment options. In this report, the successful management of a broken tusk of a juvenile male Asian elephant (Elephas maximus) including morphological analysis of the tusk tip 2 years after surgery are presented. Treatment was carried out under barn conditions and included antimicrobial photodynamic therapy and partial pulpotomy with direct pulp capping. Immediate pain relief was reached. The fractured tusk was preserved and continued to grow. The therapeutic filling material remained intact for over 1 year but was absent 2 years after treatment. The former pulp cavity of the tusk tip was filled with reparative dentin, osteodentin, and bone, but the seal between these hard tissues and pulp chamber dentin was incomplete. Radiographs obtained 3 years after treatment showed no differences in pulp shape, pulp width, and secondary dentin formation between the treated right and the healthy left tusk. It can be concluded that in case of an emergency, the endodontic therapy of a broken elephant tusk can be attempted under improvised conditions with adequate success. Photodynamic therapy might contribute to prevent infection and inflammation of the pulp. The decision tree published by Steenkamp (2019) provides a valuable tool to make quick decisions regarding a suitable therapy of broken tusks.

The Role of Dendritic Cells during Physiological and Pathological Dentinogenesis

The dental pulp is a soft connective tissue of ectomesenchymal origin that harbors distinct cell populations, capable of interacting with each other to maintain the vitality of the tooth. After tooth injuries, a sequence of complex biological events takes place in the pulpal tissue to restore its homeostasis. The pulpal response begins with establishing an inflammatory reaction that leads to the formation of a matrix of reactionary or reparative dentin, according to the nature of the exogenous stimuli. Using several in vivo designs, antigen-presenting cells, including macrophages and dendritic cells (DCs), are identified in the pulpal tissue before tertiary dentin deposition under the afflicted area. However, the precise nature of this phenomenon and its relationship to inherent pulp cells are not yet clarified. This literature review aims to discuss the role of pulpal DCs and their relationship to progenitor/stem cells, odontoblasts or odontoblast-like cells, and other immunocompetent cells during physiological and pathological dentinogenesis. The concept of “dentin-pulp immunology” is proposed for understanding the crosstalk among these cell types after tooth injuries, and the possibility of immune-based therapies is introduced to accelerate pulpal healing after exogenous stimuli.

Comparision of effectiveness of calcium hyroxide and MTA when used as an indirect pulp therapy (IPT) material- A clinical and radiological assessment

: The present study assessed and compared the success of an IPT procedure both clinically and radiographically when Dycal and MTA were used as an IPT material on primary molars.Children aged 4-9 years were screened and those who fulfilled the inclusion criteria were selected. Accordingly fiftychildren were divided into 2 groups with 25 patient in each group. Cavity preparation was done and the two test materials (Dycal and MTA) were placed at the base in their respective groups and restored with RMGIC. Post-operative radiograph was taken for baseline data. Patients were assessed at Subsequent at 1 and 6 months both clinically and radiographically. Both the test materials had formed a good biological seal, arrested further caries progression and did not cause any adverse pulpal reaction. However the amount of reparative dentin formed was highest in the Dycal group followed by MTA group. Both the experimental materials Dycal and MTA showed reparative dentin formation at the end of 1 and 6 months and also formed a good biological seal and maintained vitality of the pulp which indicates both are good IPT material.

Lithium Chloride Exerts Differential Effects on Dentinogenesis and Osteogenesis in Primary Pulp Cultures

Wnt/β-catenin signaling is known to play essential roles in odontoblast differentiation and reparative dentin formation. Various Wnt activators including LiCl have been increasingly studied for their effectiveness to induce repair of the dentin-pulp complex. LiCl is a simple salt thought to activate Wnt/β-catenin signaling by inhibiting GSK3β. Previous in vitro and in vivo studies showed that LiCl increased odontoblast differentiation and enhanced reparative dentin formation. However, the underlying molecular and cellular mechanisms by which LiCl regulates odontoblast and osteoblast differentiation during reparative dentinogenesis are not well-understood. Our in vitro studies show that exposure of early dental pulp progenitors to LiCl increased the survival and the pool of αSMA+ progenitors, leading to enhanced odontoblast and osteoblast differentiation. The positive effects of LiCl in the differentiation of osteoblasts and odontoblasts from αSMA+ progenitors are mediated by Wnt/β-catenin signaling. Our results also showed that continuous and late exposure of dental pulp cells to LiCl increased the expression of odontoblast markers through Wnt/β-catenin signaling, and the number of odontoblasts expressing DMP1-Cherry and DSPP-Cerulean transgenes. However, unlike the early treatment, both continuous and late treatments decreased the expression of Bsp and the expression of BSP-GFPtpz transgene. These observations suggest that prolonged treatment with LiCl in more mature cells of the dental pulp has an inhibitory effect on osteoblast differentiation. The inhibitory effects of LiCl on osteogenesis and Bsp were not mediated through Wnt/β-catenin signaling. These observations suggest that the effects of LiCl, and GSK3β antagonists on reparative dentinogenesis involve multiple pathways and are not specific to Wnt/β-catenin signaling.