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Ridge preservation is an important technique for maintaining the dimensions of the alveolar bone following tooth extraction, which is crucial for successful tooth rehabilitation. The combination of bovine amniotic membrane and hydroxyapatite has shown promise as a scaffold material containing growth factors that can stimulate osteogenic‐related factors such as bone morphogenetic protein 2 (BMP2), Runt‐related transcription factor 2 (RUNX2), and osteocalcin. This stimulation leads to collagen production and osteoblast proliferation, resulting in new bone formation. In this study, bovine amniotic membrane‐hydroxyapatite (BAM‐HA) composites were prepared using three different ratios of bovine amniotic membrane and hydroxyapatite (2 : 3, 3 : 7, 7 : 13). Thirty Sprague–Dawley rats had their first incisors extracted, and different types of BAM‐HA were applied for ridge preservation. The control group received no treatment, while the positive control group was given xenograft. After 14 and 28 days, the animals were sacrificed, and immunohistochemical analysis was performed to evaluate the expression of BMP2, RUNX2, and osteocalcin. Additionally, a histological examination was conducted to analyse collagen thickness and osteoblast cell proliferation. The results demonstrated that the application of BAM‐HA significantly increased collagen density, osteoblast cell proliferation, and the expression of BMP2, RUNX2, and osteoclacin compared to the control group (p < 0.05) on both days 14 and 28. Furthermore, increasing the hydroxyapatite content in the composite was found to enhance collagen thickness, osteoblast cell proliferation, and the expression of osteogenic‐related factors. These preliminary findings suggest that the combination of BAM‐HA can be used for ridge preservation to prevent further bone resorption following tooth extraction. [ABSTRACT FROM AUTHOR]

The fabrication of bio-composite-derived bovine amniotic membrane (BAM) with hydroxyapatite (HAp) is an approach to combining organic and inorganic bio-material to improve the properties of both materials. This research aims to combine, fabricate and characterise the bio-composite of BAM–HA. The combination of bio-composite is made from BAM and HAp in a ratio of 30:70, 35:65, and 40:60. Dried BAM is immersed in saline and then blended until it forms an amniotic slurry with a jelly-like consistency. At this stage, HAp is added so that it can bind to BAM. After the mixture is homogeneous, the freeze-drying process is carried out. After fabrication, all the bio-composites were characterised using Fourier transform infrared spectrometry (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM) and porosity analysis, and biological activity was conducted using fibroblasts. The bio-composite has functional groups of amides I, II, III, A, B, OH, CO32− and PO43− according to the results of the FTIR. The XRD analysis showed the presence of HAP crystals. This functional group and the crystal HAP indicate that these two materials are bound. An SEM examination revealed a variety of porous patterns on the surface area. The bio-composite with BAM and HAp at a ratio of 35:65 has a higher mean pore size of 155.625 µm with mean porosity of 89.23% and can maintain the fibroblast viability of 95.14%. In conclusion, the study successfully combined both bio-materials BAM and HAp, which have potential synergistic effects on soft and hard tissue regeneration. The ratio of 35:65 showed good characteristics and was non-toxic. [ABSTRACT FROM AUTHOR]

An active substance that has the greatest effect on wound healing is Secretory Leukocyte Protease Inhibitor (SLPI). It is known that the SLPI encoding genes can be isolated and expressed on amnion membrane. Previous studies, we isolated and optimized the SLPI gene through Echerrchia coli BL21 (DE3) mediated pET101/DTOPO, which expressed active recombinant human SLPI (rhSLPI) stored in pET-ESLPI. However, the expression of the rhSLPI products has not yet been accomplished. In this study, we optimized SLPI expression by developing a synthetic SLPI gene based on amino acid sequences with codons and expressed in E. coli BL21 to give the maximum expression. We used pUC57 and pET-32a plasmids to promote the cloning of synthetic SLPI genes. A codon)optimized SLPI gene was successfully synthesized with codon adaptation index value showing the distribution of codon usage frequency along the length of the gene sequence. In addition, the pET-SLPIopt fusion protein was successfully optimized with band sizes of 5900bp (pET-32a) and 413bp (SLPI) by double-digestion of NcoI and EcoI restriction enzymes. After the pET-SLPIopt was induced with various IPTG concentrations (50, 100, and 500 uM) at 30 °C, both soluble and insoluble fractions were analyzed as a result of SDS-PAGE which showed that the fusion protein, expressed predominantly in the supernatant, was 29.18 kDa. Our reported findings the recombinant protein of SLPI through pET-32a plasmid could be expressed in dissolved form. [ABSTRACT FROM AUTHOR]

One of dental treatments in dentistry is tooth extraction. This treatment will cause injury after extraction and biomaterial is needed to accelerate wound healing. Bovine amniotic membrane is a biomaterial with collagen content that can accelerate wound healing. However, in the process of application, the membrane will tear easily and is difficult to apply. The purpose of this study was to fabricate the bovine amniotic membrane into bovine sponge amnion (BSA) and characterize it. BSA was derived from bovine amniotic membrane with the addition of gelatin which then underwent freeze-drying process and lyophilization. BSA was then characterized using Fourier Transform Infrared Spectroscopy (FTIR), scanning electron microscopy (SEM), and swelling test. The result of characterization using FTIR indicated that amide A, amide B, amide I, amide II, and triple helix of collagen groups were shown in the FTIR graphs. The surface characterization using SEM showed a mean porosity size of 304.866 µm with a porosity proportion of 64%. The swelling analysis showed the proportion of swelling was 1320 times. The BSA has an ideal pore size, functional group and swelling ability for biomaterial candidate in dentistry. [ABSTRACT FROM AUTHOR]

Collagen sponge are used as biomaterial in the field of tissue engineering to increase the life expectancy. Bovine amniotic membrane (BAM) contains collagen and growth factors that will accelerate wound healing after dental extraction procedure. The physical structure of the BAM in the form of a thin sheet will be difficult when applied to the extraction socket. The aims of this study were to modify the physical structure of BAM in the form of a thin sheet into a sponge after that analyze its characteristics by SEM and FTIR. Fresh BAM was washed and divided into 2 parts. The first part in the form of sheets is immediately freeze dried. The second part was cut into pieces then added PBS in a ratio of 1:1. BAM and PBS are then blended until they form a slurry and are sticky like jelly, then freeze drying. After fabrication BAM and modified BAM, then it characterized by FTIR and SEM. The result of fabrication BAM looks like a thin sheet, while the modified BAM looks like a sponge that has a porous structure by using SEM analysis. FTIR analysis showed the collagen content in the presence of functional groups amide A, Amide B, amide I, amide II, amide III in BAM and modified BAM. In this study, modification of BAM will form a porous physical structure like a sponge without losing its collagen as main component. [ABSTRACT FROM AUTHOR]

Various biomaterials have been developed to be used as bone substitutes or materials to stimulate bone growth such as beta Tricalcium phosphate (beta-TCP). In addition to the selection of biocompatible materials, any technological developments have led to the project of making substitute materials leading to the manufacture of nano-sized materials. The combination of biomaterials types and size modification aims to obtain materials with biodegradability and biocompatibility abilities. The purpose of this scooping review was to get the latest description of beta tri calcium phosphate nanoparticles as scaffold combination material to increase bone formation. To obtain the data, a search was conducted using three databases, they were PubMed, Science Direct and Scopus. From all three databases, 259 articles were found. Data were then selected based on the inclusion and exclusion criteria. The title, abstract and full text of each study were carefully analyzed. Data extraction was made based on cell proliferation, cell toxicity, and new bone formation process. There were 33 selected articles and the results displayed that there were 15 other combinations of biomaterials used with beta-TCP. The in vitro studies revealed the increase in cell proliferation and the absence of toxicity from the combination of beta-TCP/hydroxyapatite and beta-TCP/Platelet Rich Plasma. While in vivo studies revealed the new bone formation from the combination of beta-TCP/hydroxyapatite and beta-TCP/ gelatin. In conclusion, the comparison of ingredients combined with beta-TCP in this scooping review were hydroxyapatite, platelet rich plasma and gelatin. Various combinations of these biomaterials with beta-TCP had high cell proliferation ability, no cell toxicity and good new bone formation ability. [ABSTRACT FROM AUTHOR]

Bovine sponge amnion (BSA) is made from bovine amniotic membrane, which contains several growth factors with the ability to suppress inflammation and stimulate the healing process. The healing process of the socket is clinically marked by the proliferation of fibroblasts, formation of new blood vessels, and closure of the wound site. Fibroblasts and new blood vessels are the common histological indicators of tissue regeneration. Twenty-four lower anterior teeth were extracted from the rats. The socket was then applied with BSA and sutured, while a control group was only sutured. The animals were scarified 3, 7, and 14 days after application, and the mandibular was isolated. Histological analysis of socket tissue used the staining of hematoxylin and eosin to analyze the new blood vessels and fibroblasts. An independent t-test was used to analyze the fibroblasts and new blood vessels in each group, with P < 0.05 considered as significant. The number of fibroblasts is higher in the BSA group compared others groups at 3, 7, and 14 days (P < 0.05). The new blood vessel count is higher compared to the control group at only 3 and 7 days (P < 0.05). BSA aids the regeneration of the socket after tooth extraction by stimulating fibroblast proliferation and formation of new blood vessels. [ABSTRACT FROM AUTHOR]

The secretory Leukocyte Protease Inhibitor (SLPI) is a protease inhibitor which can be found in saliva, bronchial mucus, seminal plasma and the amniotic membrane. It has been recognized for anti-inflammatory, antimicrobial and wound healing. Recombinant SLPI produced in Saccharomyces cerevisiae would be a useful biomaterial for wound healing. The yeast as a heterologous host holds an important role to express human SLPI. The yeast PDI is an enzyme which is required in the formation of disulfide bonds and folding of SLPI. A co-expression vector harboring SLPI gene and PDI1 gene is constructed aiming to strengthen the expression of SLPI in S. cerevisiae. The SLPI gene from the amniotic membrane is fused with HM1 signal peptide, namely hmSLPI. The HM1 signal peptide is used to mediate the translocation of SLPI across the membrane cells. The plasmids construction and characterization are done in Escherichia coli TOP10 prior to its transformation to S. cerevisiae BJ1824. The hmSLPI fusion gene and PDI1 gene are successfully amplified in sized 483 bp and 1,583 bp, respectively. The restriction analysis and nu- cleotide sequencing indicate that the recombinant plasmids, pAT_hmSLPI, pAT_PDI1 and pAT_PDI1_hmSLPI, are successfully constructed. All of the recombinant plasmids are also successfully introduced into S. cerevisiae BJ1824. [ABSTRACT FROM AUTHOR]

Alveolar bone resorption will occur in 40--60% within three months after tooth extraction. Socket preservation is performed to reduce the occurrence of alveolar bone resorption for successful implant restoration. One of the successes of implant restoration is in the quality and quantity of alveolar bone. The combination of the bovine amniotic membrane and hydroxyapatite is expected to be a candidate for socket preservation material. This review article aimed at explaining the potential of bovine amniotic membrane and hydroxyapatite that can act as biocomposite materials for socket preservation to enhance bone formation. [ABSTRACT FROM AUTHOR]

Background: Wound tooth extraction is a mechanical injury that traumatizes adjacent tissue. Sponge amnion contains growth factors that can promote postextraction wound healing. Amnion membranes can be transformed into sponge form rendering it easier to use. The aim of this study is to analyze interleukin-6 (IL-6) and bone morphogenetic protein-2 (BMP-2) expression in postextraction wound healing on the 1st and 7th day after sponge amnion application. Materials and Methods: Twenty-eight Wistar rats were used in this experimental descriptive analytical study. Fourteen animals' first right anterior mandible tooth was extracted; then, the socket applied by sponge amnion and sutured (treatment group), while 14 others only sutured (as control group). The alveolar bone tissue of animal was observed 1st and 7th days after extraction and then was analyzed using immunohistostaining to identify the expression of IL-6 and BMP-2. Statistical analysis was performed using one-way ANOVA with the level of significance (P < 0.05). Results: IL-6 expression in the treatment group was significantly lower than the control group on the 1st and 7th days (P = 0.000). BMP-2 expression in the treatment group was significantly higher than the control group on the 1st and 7th days (P = 0.000). Conclusion: Sponge amnion can promote the healing process by increasing the expression of BMP-2 and decreasing IL-6 expression. [ABSTRACT FROM AUTHOR]

Biomaterials that can be used to accelerate wound healing is the amniotic membrane that contains growth hormone. In its use, amniotic membrane tends to be difficult to use because of their sheets form. To make it easier to use, the amniotic membrane is mixed with gelatin as an adhesive material to produce sponge amnion. Sponge amnion which is used as a biomaterial needs to meet several requirements, one of them is biocompatibility. To complete these requirements cytotoxicity testing is necessary to determine the toxic potential that a material may produce. The cytotoxicity test was performed on BHK-21 fibroblast cells. The aim of this study is to determine the citotoxicity effect of amnion in sponge form on BHK-21 fibroblast cells. This study was done towards the culture cell line BHK-21 using MTT assay method. In this study used four sample groups. Two groups are the control group (media control and cell control), and the other two groups are samples consisted of amniotic membrane and sponge amnion. Living cells were quantified after treatment by ELISA reader on 620 nm. The fibroblast cell life percentage after exposure with amniotic membrane and sponge amnion is 98,16% and 98.13%. Based on the ANOVA test result was obtained P-value of 0.000 (P-value <0.05) so that it can be said that the treatment result gives a significant difference. Amnion in sponge form has no toxic for BHK-21 fibroblast cell. [ABSTRACT FROM AUTHOR]

Amniotic membrane is one of the natural biomaterial that is widely used for wound healing which has Secretory Leukocyte Protease Inhibitor (SLPI) as an active component. One of rSLPI functions is suppressing several pro-inflammatory factors, such as serine protease. Supression of serine proteinase in wound healing process may increase the amount of vascular endothelial growth factor (VEGF) and collagen. The aim of this study is to evaluate the effect of rSLP amniotic membrane fluid application on incision wound of Rattus norvegicus in expression of VEGF and collagen. The rats divided into 4 groups: control group and treated group with rSLPI (P1 0.03 cc, P2 0,045 cc, and P3 0.06 cc) (n=6). Insicion was made on rat's cervical back. On the day 4, tissues around insicion wound are collected for Masson's Trychrome (MT) and Immunohistochemistry (IHC) analysis. Expression of VEGF and collagen are calculated using microscope with 400x magnification. Kruskal-Wallis test of VEGF showed significant value with p=0,006 (p<0.05). Mann-Whitney test of VEGF showed a significant difference between control and treatment group. Oneway ANOVA test result for collagen has p value 0.000 (p<0.05). Honestly Significant Dif ference (HSD) test showed a significant differences between all group. Application of rSLPI amniotic membran fluid in Rattus norvegicus incision wound may increase VEGF and collagen. [ABSTRACT FROM AUTHOR]

Bone defect is still a major problem in dentistry. In oral cavity, trauma, congenital disorder, periodontitis and tumor can damage bones. A biomimetic scaffold made of bovine hydroxyapatite (BHA), gelatin (GEL), and chitosan (K) (BHA-GEL-K) is on a development in recent researches. This research tends to develop BHA-GEL-K scaffolds combined with 0.25% glutaraldehyde as cross linking agent. Scaffold's profiles were examined with FTIR, porosity size by using SEM and compressive strength by using autograph. The adhesion of bone narrow mesenchymal stem cell (BM-MSC) to crosslinked BHA-GEL-K scaffolds with 0.25% glutaraldehyde were examined by using SEM. BHA, GEL, K and glutaraldehyde components are bonded to each other. These scaffolds have 121.21 μm of porosity and 912.86 + 83.81 kPa of compressive strength value. These scaffolds have good adhesion to BM-MSC. The adhesion of scaffolds increases since day 1 to 7. The profile of BHA-GEL-K scaffolds that crosslinked to 0.25% glutaraldehyde may fulfills a requirement of bone tissue engineering. [ABSTRACT FROM AUTHOR]

Background: The Secretory Leukocyte Protease Inhibitors (SLPI) has many biological functions including anti-bacterial, anti-fungal, anti-viral, anti-inflammatory, and immuno-modulatory. Previous studies have shown that gene-encoding human SLPI have successfully been expressed in Escherichia coli (E. coli) with a C-terminal poly-histidine tag (His-tag). The aim of this research was to investigate the inhibition activity of N-terminal His-tag position (NSLPI) and C-terminal His-tag position (CSLPI). We hypothesized that a His-tag close to an active site SLPI domain may interfere with the inhibition activity of SLPIs., Methods: A NSLPI and CSLPI were constructed with polymerase chain reaction (PCR) amplification. The PCR products were then ligated into pET-30a plasmid and transformed into E. coli TOP10. Recombinant plasmids were verified by using restriction analysis and nucleotide sequence analysis. pET-NSLPI and pET-CSLPI were then subcloned in E. coli BL21(DE3) for its expression. The SLPI protein was expressed using Isopropyl β-D-1-thiogalactopyranoside induction (IPTG). The inhibition effect of both SLPI against Porcine Pancreatic Elastase (PPE) enzyme was tested using the N-succinyil-alanyl-L-alanyl-L-prolyl-L-phenylalanyl-4-nitroanalide (NPN) substrate., Results: The SLPI gene was successfully cloned and expressed in E. coli BL21. Fusion proteins of NSLPI and CSLPI were generated with His-tag in the N-terminal and C-terminal position, respectively. The inhibition effect of NSLPI and CSLPI on PPE indicated that both SLPI were active. The inhibition activity of NSLPI was 66.7%, higher than CSLPI by 44.4%., Conclusion: The His-tag position on the C-terminal of SLPI reduced the inhibition activity of SLPI., Competing Interests: Conflict of Interest The authors declare that there are no conflicts of interest regarding the publication of this paper., (Copyright© 2020 Avicenna Research Institute.)