172
Dental Journal
(Majalah Kedokteran Gigi)
2023 September; 56(3): 172–177
Original article
The characteristics of swelling and biodegradation tests of
bovine amniotic membrane-hydroxyapatite biocomposite
Titien Hary Agustantina
1
, Elly Munadziroh
1
, Anita Yuliati
1
, Muhammad Riza Hafidz Bahtiar
2
, Octarina
3,4
, Rizki Fauziah Salma
2
, Ajeng Putri
Meyranti
2
, Fathilah Abdul Razak
5
1
Department of Dental Materials, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia
2
Undergraduate Student, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia
3
Doctoral Program, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia
4
Department of Dental Materials, Faculty of Dentistry, Universitas Trisakti, Jakarta, Indonesia
5
Department of Oral and Craniofacial Sciences, Faculty of Dentistry, Universiti Malaya, Kuala Lumpur, Malaysia
ABSTRACT
Background: A good biocomposite is a structure that can provide opportunities for cells to adhere, proliferate, and differentiate.
It is affected by the characteristics of a material. As bone tissue regeneration occurs, biomaterials must have a high swelling ability
and low biodegradability. The high swelling capability will have a larger surface area that can support maximal cell attachment and
proliferation on the biocomposite surface, which accelerates the regeneration process of bone defects.
Purpose: The study aimed to
analyze the characteristics of swelling and biodegradation of bovine amniotic membrane-hydroxyapatite (BAM-HA) biocomposite with
various ratios. Methods: The BAM-HA biocomposite with a ratio of 30:70, 35:65, and 40:60 (w/w) was synthesized using a freeze-dry
method. The swelling test was done by measuring the initial weight and final weight after being soaked in phosphate-buffered saline
for 24 hours and the biodegradation test was done by measuring the initial weight and final weight after being soaked in simulated
body fluid for seven days. Results: The swelling percentage of BAM-HA biocomposite at each ratio of 30:70, 35:65, and 40:60 (w/w)
was 303.90%, 477.94%, and 574.19%. The biodegradation percentage of BAM-HA biocomposite at each ratio of 30:70, 35:65, and
40:60 was 9.43%, 11.05%, and 12.02%. Conclusion: The BAM-HA biocomposite with a ratio of 40:60 (w/w) has the highest swelling
percentage while the 30:70 (w/w) ratio has the lowest percentage of biodegradation.
Keywords: biocomposite; biodegradation; bovine amniotic membrane; hydroxyapatite; socket preservation; swelling
Article history: Received 10 October 2022; Revised 23 November 2022; Accepted 12 January 2023; Published 1 September 2023
Correspondence: Elly Munadziroh, Department of Dental Materials, Faculty of Dental Medicine, Universitas Airlangga. Jl. Mayjen
Prof. Dr. Moestopo 47 Surabaya, 60132, Indonesia. Email: [email protected]
INTRODUCTION
The desired outcomes of biomaterials related to tissue
engineering for clinical applications should include the
attraction of natural resident stem cells to the site of
injury and the suppression of inflammation, reduction
of scar formation, enhancement of vascularization, and
prevention of infection. The amniotic membrane is
recommended for usage as an ideal biomaterial for wound
healing due to its protein, cytokine, and growth factor
concentration. The amniotic membrane can be used as a
single or even double sheet to cover wounds on the body’s
exposed outer surfaces, such as the skin and cornea.
1
The amniotic membrane has many different applications
in medicine, but it is particularly useful for treating skin
burns and preventing tissue adhesion during head, neck,
abdomen, larynx, and genitourinary tract surgeries. The
amniotic membrane can also serve as a barrier that protects
internal organs.
2
For instance, wrapping the peritoneal
cavity, tendon, spinal cord, and peripheral nerves in the
amniotic membrane could prevent adhesion and minimize
the formation of scars.
1
Amniotic membrane is a biomaterial used widely
in tissue regeneration because it has anti-bacterial and
anti-inflammatory properties.
3
Amniotic membrane has
growth factors, such as vascular endothelial growth factor
(VEGF), fibroblast growth factor (FGF), epidermal growth
factor (EGF), tissue inhibitor metalloproteinase (TIMP),
transforming growth factor-β (TGF-β), and platelet-
derived growth factor (PDGF).
4
One source of the amniotic
Copyright © 2023 Dental Journal (Majalah Kedokteran Gigi) p-ISSN: 1978-3728; e-ISSN: 2442-9740. Accredited No. 158/E/KPT/2021.
Open access under CC-BY-SA license. Available at https://e-journal.unair.ac.id/MKG/index
DOI: 10.20473/j.djmkg.v56.i3.p172–177
173Agustantina et al. Dent. J. (Majalah Kedokteran Gigi) 2023 September; 56(3): 172–177
membrane is bovine. The bovine amniotic membrane has
some benefits over the human amniotic membrane, such
as being easier to legalize, being considered more ethical,
and not being impacted by religious rituals.
5
The limitation of the amniotic membrane is its low
mechanical properties and rapid degradation, which makes
it easy to decompose and difficult to maintain the structural
integrity required for bone regeneration.
6
The amniotic
membrane is also easily torn by this feature when it is
applied. The integrity of the structure, which is vital when
mending extraction wounds with long-term treatment,
will be impacted by how easily the amniotic membrane
degrades. Modifying the amniotic membrane is needed to
find a solution to this issue.
7
The amniotic membrane in
combination with a bone graft will produce biocomposites
that have the potential to support bone formation and
provide better outcomes.
8
One of the bone graft materials that is often used to support
the bone healing process is hydroxyapatite.
9
Hydroxyapatite
is a bioceramic material with a mineral composition
similar to bones and teeth.
10,11
Hydroxyapatite has been
used widely as a biomaterial for bone tissue replacement
and repair because of its high osteoconductivity, non-
toxicity, and good biocompatibility.
12
Hydroxyapatite can
be obtained from bovine bone (bovine hydroxyapatite).
13
Bovine bone material consists of 93% hydroxyapatite and
7% β-tricalcium phosphate (Ca
3(PO
4)
2, β-TCP).
14
Bovine
hydroxyapatite (BHA) has the same chemical composition
as human bone, which affects bone metabolism.
15
A combination of bovine amniotic membrane and
hydroxyapatite will produce a new material in the form
of a sponge-shaped biocomposite. The bovine amnion-
hydroxyapatite membrane was expected to preserve the
socket after tooth extraction. The characteristic properties
of a biomaterial are important for successful tissue
regeneration and must be able to adapt to the tissue that
will be replaced.
16
A good biocomposite is a structure that
can provide opportunities for cells to adhere, proliferate,
and differentiate. It is affected by the characteristics of a
material.
17
The swelling test is a method used to determine the
capacity of the material to absorb liquid. The swelling
properties of the biomaterial have an impact on good cell
proliferation. Higher swelling abilities increase the surface
area of the biomaterial, thus facilitating cell attachment
to the biomaterial.
18
With more cells attached, it is hoped
that cell growth will also be faster. Swelling properties
also play an important role in increasing the absorption
of fluids from the body or media, as well as the transfer
of nutrients and metabolic wastes.
19
The increase in the
swelling ratio is caused by the presence of hydrophilic
properties. Hydrophilic conditions are suitable for cell
attachment and proliferation, therefore it accelerates the
regeneration process.
20
Biomaterials implanted in the body and in contact
with biological systems can trigger a series of reactions
between the biomaterials and host tissues. The ability
of biodegradation is an important role in biomaterials’
formation of new tissues because of its properties that can
affect cell viability and proliferation. The biodegradation
test is a parameter needed to see the time required for
biomaterial to be degraded according to the formation
of new tissue. The biodegradation test can indicate the
biodegradability of a material.
21
Biomaterials that have been implemented in the body
must be able to maintain sufficient mechanical properties
and structural integrity so that cell adaptation goes well and
can store their extracellular matrix. Biomaterials that have
biodegradable properties are expected to be able to create
space for new bone tissue to grow.
22
Based on this, the study
aimed to analyze the characteristics of bovine amniotic
membrane-hydroxyapatite (BAM-HA) biocomposite
through a swelling and biodegradation test.
MATERIALS AND METHODS
Ethical approval, which is managed as a condition for
conducting research, has been obtained from the Research
Ethics Commission of the Faculty of Medicine, Universitas
Airlangga Surabaya (No. 400/HRECC.FODM/VII/2021).
This research is an experimental laboratory with a post-
test-only control group design. The data used primary data,
which was directly collected by the researcher.
The manufacture of BAM-HA biocomposite was done
at the Biomaterial Center Installation of the Tissue Bank
Hospital by Dr. Soetomo Surabaya. The dry amniotic
membrane was prepared at 3 grams (30:70 ratio), 3.5 grams
(ratio 35:65), and 4 grams (ratio 40:60). The amniotic
membrane was cut into pieces of about 2 cm and added
with 40 ml of 0.9% NaCl. Then, the amniotic membrane
with NaCl was soaked for 5 minutes until the liquid was
absorbed. The amniotic membrane was blended for 10
minutes to produce a smooth amniotic slurry. The amniotic
slurry was added with 7 grams of hydroxyapatite powder
(30:70 ratio), 6.5 grams of hydroxyapatite powder (ratio
35:65), and 6 grams of hydroxyapatite powder (ratio 40:60),
then stirred until homogeneous and put into a petri dish with
a diameter of 10 cm. The petri dish was put into the freezer
(Thermo, USA) at -80
o
C for 24 hours, then freeze-dried
(VirTis BenchTop
TM
“K” Series) for 24 hours at -100
o
C
(Figure 1). Sponge-shaped biocomposite with a diameter
of 10 cm was cut into pieces of 1.5x1.5 cm using a scalpel
with a no. 15 blade and handle scalpel no. 3 (Figure 2).
The biocomposite is in the form of a sponge with a
sizing of 1.5x1.5 cm and various ratios weighed as the
initial weight (Wi). The biocomposite was immersed in 10
ml of Phosphate Buffer Saline (PBS) solution at 37
o
C for
24 hours. After that, the biocomposite was taken and then
drained using Whatman filter paper for 3 seconds, and the
final weight (Wf) was determined. The swelling ratio was
calculated using the formula:
Copyright © 2023 Dental Journal (Majalah Kedokteran Gigi) p-ISSN: 1978-3728; e-ISSN: 2442-9740. Accredited No. 158/E/KPT/2021.
Open access under CC-BY-SA license. Available at https://e-journal.unair.ac.id/MKG/index
DOI: 10.20473/j.djmkg.v56.i3.p172–177
174 Agustantina et al. Dent. J. (Majalah Kedokteran Gigi) 2023 September; 56(3): 172–177
Table 1.
The average value of the swelling test
Biocomposite Ratio
Swelling ratio (%)
Mean SD
Bovine amniotic membrane-
hydroxyapatite
30:70 (Control) 303.90 10.75
35:65 477.94 10.90
40:60 574.19 35.44
Table 2.
The results of one-way Welch ANOVA statistical analysis on the swelling test
Robust tests of equality of means
Swelling test Statistic df1 df2 Sig.
Welch 745.760 2 16.455 .000
Table 3.
Post-hoc Tukey HSD statistical test results of swelling percentage
Ratio 30:70 35:65 40:60
30:70 0.000* 0.000*
35:65 0.000*
40:60
*Significant (p < 0.05)
A B
C
Figure 1.
The results of making BAM-HA ratios of 30:70 (A), 35:65 (B), and 40:60 (C).
A B C
Figure 2.
The results of the preparation of BAM-HA biocomposite ratios of 30:70 (A), 35:65 (B), and 40:60 (C).
The biodegradation test was performed using a
biocomposite is in the form of a sponge with a sizing of
1.5x1.5 cm and various ratios weighed as the Wi. The
biocomposite was immersed in Simulated Body Fluid
(SBF) solution at 37°C for seven days. After seven days,
the biocomposite was taken and then dried using a freeze-
drier for 48 hours at a temperature of -100
o
C. After that,
the biocomposite was weighed again to determine the Wf.
The biodegradation of biocomposites was calculated using
the formula:
RESULTS
The average value of the swelling level of the BAM-HA
biocomposite (Table 1) increased after immersion for 24
hours. The BAM-HA biocomposite with a ratio of 40:60
(w/w) obtained an average swelling rate of 574.19%,
which was the highest rate compared to the ratios of 35:65
and 30:70 (w/w), which were 477.94% and 303.90%,
respectively.
Statistical analysis of variance (one-way Welch
ANOVA) is one of the parametric statistical tests to
Copyright © 2023 Dental Journal (Majalah Kedokteran Gigi) p-ISSN: 1978-3728; e-ISSN: 2442-9740. Accredited No. 158/E/KPT/2021.
Open access under CC-BY-SA license. Available at https://e-journal.unair.ac.id/MKG/index
DOI: 10.20473/j.djmkg.v56.i3.p172–177
175Agustantina et al. Dent. J. (Majalah Kedokteran Gigi) 2023 September; 56(3): 172–177
determine differences between groups of data as a whole.
The results of the one-way Welch ANOVA test showed
a significance value of p = 0.000, which means that
there was a significant difference between groups with a
p-value of less than (p < 0.05) (Table 2). Therefore, further
testing was carried out using the post-hoc Tukey Honestly
Significant Difference (HSD) to find out which groups had
significant differences. The post-hoc Tukey HSD test has
been summarized and presented in Table 3.
There was a significant difference in the average level
of swelling between the BAM-HA biocomposite ratio of
30:70 with a ratio of 35:65 (w/w) (p = 0.000), BAM-HA
biocomposite ratio of 30:70 with a ratio of 40:60 (w/w) (p
= 0.000), and the BAM-HA biocomposite ratio of 35:65
with a ratio of 40:60 (w/w) (p = 0.000) (p < 0.05).
The average value of the biodegradation rate of BAM-
HA biocomposite in various ratios is shown in Table 4.
The BAM-HA biocomposite 40:60 (w/w) ratio obtained
the highest average value of the biodegradation rate at
12.02%, followed by 11.05% at 35:65 (w/w), and 9.54%
at 30:70 (w/w).
The results of the one-way Welch ANOVA test showed
a significance value of p = 0.000, which means that there
was a significant difference between groups with a p-value
of less than 0.05 (Table 5). Therefore, the post-hoc Tukey
HSD analysis test was continued to find out which groups
had significant differences. The post-hoc Tukey HSD test
has been summarized and presented in Table 6.
Data analysis was performed using the post-hoc
Tukey HSD test. It showed that there was a significant
difference in the mean level of biodegradation between the
BAM-HA biocomposite group with a ratio of 30:70 and a
comparison ratio of 35:65 (w/w) (p = 0.000) and BAM-HA
biocomposite with a ratio of 30:70 and a comparison ratio
of 40:60 (w/w) (p = 0.000).
DISCUSSION
The selection of biomaterials for making bone tissue
regeneration material is a significant matter in the socket
preservation procedure. The requirements for biomaterials
in bone tissue regeneration, include having a porous
structure that promotes cell proliferation and distribution
so that it can support the formation of new bone tissue.
Additionally, it can provide a temporary structure that will
degrade over time with the formation of new tissue.
23,24
Socket preservation biomaterial candidates used in
this study were obtained by synthesizing bovine amniotic
membrane biomaterials and BHA with a combined ratio
of 30:70, 35:65, and 40:60 (w/w) using the freeze-drying
method. The freeze-drying process can form a porous
structure, which can affect the outer surface of the
biocomposite to become hydrophilic.
25,26
The characteristic properties of a biomaterial are
important for the success of tissue regeneration and must be
able to adapt to the tissue to be replaced.
16
Swelling is one
of the important properties in the application of biomaterials
in bone tissue regeneration. Swelling in biomaterials
can play a role in cell infiltration into biomaterials.
27
The results of the swelling test showed that the average
swelling ratio was increasing from a ratio of 30:70 (w/w)
of 303.90±10.75%, 35:65 (w/w) of 477.94±10.90%, 40:60
(w/w) of 574.19±35.44%, and after the one-way ANOVA
test, showed that there was a significant difference between
groups with a p-value of less than 0.05.
This study showed that the BAM-HA biocomposite
with the larger bovine amniotic membrane ratio of 40:60
(w/w) obtained the highest percentage of swelling results
compared to the 35:65 and 30:70 (w/w) ratios. That is
because the bovine amniotic membrane biomaterial in
this study contains a polymer in the form of collagen.
28
Table 4.
The average value of the swelling test
Biocomposite Ratio
Biodegradation ratio (%)
Mean SD
Bovine amniotic membrane-
hydroxyapatite
30:70 (Control) 9.54 1.43
35:65 11.05 1.42
40:60 12.02 0.48
Table 5.
The results of one-way Welch ANOVA statistical analysis on the biodegradation test
Robust Tests of Equality of Means
Biodegradation test Statistic df1 df2 Sig.
Welch 14.048 2 14.307 .000
Table 6.
Post-hoc Tukey HSD statistical test results of biodegradation percentage
Ratio 30:70 35:65 40:60
30:70 0.024* 0.000*
35:65 0.180
40:60
*Significant (p < 0.05)
Copyright © 2023 Dental Journal (Majalah Kedokteran Gigi) p-ISSN: 1978-3728; e-ISSN: 2442-9740. Accredited No. 158/E/KPT/2021.
Open access under CC-BY-SA license. Available at https://e-journal.unair.ac.id/MKG/index
DOI: 10.20473/j.djmkg.v56.i3.p172–177
176 Agustantina et al. Dent. J. (Majalah Kedokteran Gigi) 2023 September; 56(3): 172–177
These findings are appropriate with several findings from
previous research that demonstrates collagen is one of the
polymers with a high-water absorption capacity. A high
amount of collagen will increase the water absorption
properties of the biomaterial. Collagen is a polymer with
many hydrophilic groups, which can form hydrogen bonds
with the surrounding solution. The low contact angle of
biomaterials with hydrophilic characteristics can result in
low surface tension. If the surface tension of the liquid is
low, the surface of the biocomposite material will contact
it more easily. As a result, the biocomposite can absorb
liquids more readily.
29
Several other studies have shown that the increase in
the percentage of swelling is directly proportional to the
increase in the amount of gelatin, which is widely known
as a biomaterial that has hydrophilic properties, while
the increase in the amount of hydroxyapatite is inversely
proportional to the percentage of swelling. The higher
hydroxyapatite ratio in the biocomposite caused a decrease
in the percentage of swelling and its swelling ability.
19,30
By attaching calcium and phosphate to the hydrophilic
groups of COOH or NH
2, hydroxyapatite forms a cross-
link between polymer chains and reduces the biomaterial’s
hydrophilicity.
31
Another property that plays a role in the success of
bone tissue regeneration is biodegradability. Biodegradable
biomaterials were expected to provide a space for the growth
of new bone tissue.
22
According to the data, the BAM-HA
biocomposite ratio of 40:60 (w/w) had the highest average
value of biodegradation at 12.020±0.483%, followed by
11.045±1.415% at a ratio of 35:65 (w/w). Meanwhile,
the BAM-HA biocomposite with a ratio of 30:70 (w/w)
had the lowest percentage of biodegradation, which was
9.541±1.428%. The results of the one-way Welch ANOVA
test showed a value of significance of p = 0.000, which
means that there is a significant difference between groups
with a p-value of less than 0.05. The lowest percentage of
a biodegradation rate was obtained at a 30:70 (w/w) ratio,
with the highest composition hydroxyapatite ratio. That
is because hydroxyapatite can reduce the pore size of the
biocomposite.
30
As a result, the lowest percentage level
was attained at the 30:70 (w/w) ratio, which had a higher
hydroxyapatite content than the 40:60 and 35:65 (w/w)
ratios. Meanwhile, the average percentage of degradation
rate was high for the 40:60 (w/w) ratio because it contained
less hydroxyapatite and more bovine amniotic membrane
than the ratios of 35:65 and 30:70 (w/w).
The inclusion of the hydroxyapatite ratio can result
in a reduction in porosity size. Therefore, it may have
an impact on a biocomposite’s ability to access liquids.
When the biomaterial immerses in the SBF solution, the
solution forms a link with the biomaterial’s surface. By
capillary action, the fluid will progressively seep into the
biomaterial. During this phase, the pore wall will serve as
a new connection between the liquid and the biomaterial.
A decrease in porosity size will result in a smaller surface
area. As a result, liquid accessibility reduces, which results
in a slower rate of biodegradation.
32
The level of biomaterial
degradation is influenced by chemical properties, polymer
composition, and environmental conditions.
18
This study seeks to identify the appropriate combination
ratio between the bovine amniotic membrane and
hydroxyapatite and develop a bone biocomposite that
produces swelling and biodegradation properties suitable
for bone tissue regeneration. As bone tissue regeneration
occurs, biomaterials must have a high swelling ability
and low biodegradability. The high swelling capability
will have a larger surface area that can support maximal
cell attachment and proliferation on the biocomposite
surface, which accelerates the regeneration process of bone
defects.
27
Biocomposite can be used as bone graft material,
preferably having a low level of biodegradation for bone
regeneration to occur.
26
The results show that the percentage of swelling increased
with the concentrations of bovine amniotic membrane
increasing and the concentrations of hydroxyapatite
decreasing. Additionally, the percentage of biodegradation
decreased with the concentrations of hydroxyapatite
increasing and the concentrations of bovine amniotic
membrane decreasing. The BAM-HA biocomposite with a
ratio of 40:60 (w/w) has high swelling and biodegradation
ability, while the ratio of 30:70 (w/w) has low swelling and
biodegradation ability.
REFERENCES
1.
Elkhenany H, El-Derby A, Abd Elkodous M, Salah RA, Lotfy
A, El-Badri N. Applications of the amniotic membrane in tissue
engineering and regeneration: the hundred-year challenge. Stem
Cell Res Ther. 2022; 13(1): 8.
2.
Schmiedova I, Dembickaja A, Kiselakova L, Nowakova B, Slama P.
Using of amniotic membrane derivatives for the treatment of chronic
wounds. Membranes (Basel). 2021; 11(12): 941.
3.
Kumar A, Chandra RV, Reddy AA, Reddy BH, Reddy C, Naveen
A. Evaluation of clinical, antiinflammatory and antiinfective
properties of amniotic membrane used for guided tissue regeneration:
A randomized controlled trial. Dent Res J (Isfahan). 2015; 12(2):
127–35.
4.
Faadhila T, Valentina M, Munadziroh E, Nirwana I, Soekartono
H, Surboyo MC. Bovine sponge amnion stimulates socket
healing: A histological analysis. J Adv Pharm Technol Res. 2021;
12(1): 99.
5.
Putra NHD, Matulatan F, Wibowo MD, Danardono E. Effects of
dried bovine amniotic membrane as prosthetics of abdominal fascial
defect closure observed by the expression of platelet-derived growth
factor in Rattus norvegicus wistar strain. Syst Rev Pharm. 2020;
11(6): 987–91.
6.
Leal
‐Marin S, Kern T, Hofmann N, Pogozhykh O, Framme C,
Börgel M, Figueiredo C, Glasmacher B, Gryshkov O. Human
amniotic membrane: A review on tissue engineering, application,
and storage. J Biomed Mater Res Part B Appl Biomater. 2021; 109(8):
1198–215.
7.
Dadkhah Tehrani F, Firouzeh A, Shabani I, Shabani A. A review on
modifications of amniotic membrane for biomedical applications.
Front Bioeng Biotechnol. 2021; 8: 606982.
8.
Nurhaeini CSW, Komara I. Socket preservation. Padjadjaran J Dent.
2015; 27(3): 133–8.
9.
Arifin A, Mahyudin F, Edward M. The clinical and radiological
outcome of bovine hydroxyapatite (bio hydrox) as boneGraft. J
Orthop Traumatol Surabaya. 2020; 9(1): 9–16.
Copyright © 2023 Dental Journal (Majalah Kedokteran Gigi) p-ISSN: 1978-3728; e-ISSN: 2442-9740. Accredited No. 158/E/KPT/2021.
Open access under CC-BY-SA license. Available at https://e-journal.unair.ac.id/MKG/index
DOI: 10.20473/j.djmkg.v56.i3.p172–177
177Agustantina et al. Dent. J. (Majalah Kedokteran Gigi) 2023 September; 56(3): 172–177
10.
Kattimani VS, Kondaka S, Lingamaneni KP. Hydroxyapatite–-Past,
present, and future in bone regeneration. Bone Tissue Regen Insights.
2016; 7: BTRI.S36138.
11.
Rujitanapanich S, Kumpapan P, Wanjanoi P. Synthesis of
hydroxyapatite from oyster shell via precipitation. Energy Procedia.
2014; 56: 112–7.
12.
Ardhiyanto HB. Peran hidroksiapatit sebagai material bone
graft dalam menstimulasi kepadatan kolagen tipe L pada proses
penyembuhan tulang. Stomatognaic - J Kedokt Gigi. 2012; 9(1):
16–8.
13.
Yuliati A, Kartikasari N, Munadziroh E, Rianti D. The profile of
crosslinked bovine hydroxyapatite gelatin chitosan scaffolds with
0.25% glutaraldehyde. J Int Dent Med Res. 2017; 10(1): 151–5.
14.
Saputra AAH, Triyono J, Triyono T. Bovine bone hidroksiapatite
materials mechanics properties at 900°C and 1200°C of calcination
temperature. Mek Maj Ilm Mek. 2017; 16(1): 26–30.
15.
Ratnayake JTB, Mucalo M, Dias GJ. Substituted hydroxyapatites
for bone regeneration: A review of current trends. J Biomed Mater
Res Part B Appl Biomater. 2017; 105(5): 1285–99.
16.
Shoichet MS. Polymer scaffolds for biomaterials applications.
Macromolecules. 2010; 43(2): 581–91.
17.
Bru
žauskaitė I, Bironaitė D, Bagdonas E, Bernotienė E. Scaffolds and
cells for tissue regeneration: different scaffold pore sizes—different
cell effects. Cytotechnology. 2016; 68(3): 355–69.
18.
Kartikasari N, Yuliati A, Listiana I, Setijanto D, Suardita K,
Ariani MD, Sosiawan A. Characteristic of bovine hydroxyapatite-
gelatin-chitosan scaffolds as biomaterial candidate for bone tissue
engineering. In: 2016 IEEE EMBS Conference on Biomedical
Engineering and Sciences (IECBES). Kuala Lumpur, Malaysia:
IEEE; 2016. p. 623–6.
19.
Azhar FF, Olad A, Salehi R. Fabrication and characterization of
chitosan-gelatin/nanohydroxyapatite- polyaniline composite with
potential application in tissue engineering scaffolds. Des Monomers
Polym. 2014; 17(7): 654–67.
20.
Mohamed KR, Beherei HH, El-Rashidy ZM. In vitro study of nano-
hydroxyapatite/chitosan–gelatin composites for bio-applications. J
Adv Res. 2014; 5(2): 201–8.
21.
Maji K, Dasgupta S, Pramanik K, Bissoyi A. Preparation
and evaluation of gelatin-chitosan-nanobioglass 3D porous
scaffold for bone tissue engineering. Int J Biomater. 2016; 2016:
9825659.
22.
Bose S, Roy M, Bandyopadhyay A. Recent advances in bone tissue
engineering scaffolds. Trends Biotechnol. 2012; 30(10): 546–54.
23.
Melek LN. Tissue engineering in oral and maxillofacial
reconstruction. Tanta Dent J. 2015; 12(3): 211–23.
24.
Edgar L, McNamara K, Wong T, Tamburrini R, Katari R, Orlando
G. Heterogeneity of scaffold biomaterials in tissue engineering.
Materials (Basel). 2016; 9(5): 332.
25.
Tam TT, Todo M, Cheong KY, Hamid ZAA. Evaluation of cell
viability of porous scaffold fabricated via freeze-drying technique
for vascular tissue engineering. In: AIP Conference Proceedings.
2020. p. 020007.
26.
Wattanutchariya W, Changkowchai W. Characterization of porous
scaffold from chitosan-gelatin/hydroxyapatite for bone grafting.
In: The International MultiConference of Engineers and Computer
Scientists Vol II. Hongkong: Chiang Mai University; 2014.
p. 1–5.
27.
Asaeli AP, Yuliati A, Budhy TI, Wilda S, Silalahi I, Santoso D. The
profile of cross-linked chitosan and collagen derived-chicken shank
scaffold as biomaterials in tissue engineering. J Int Dent Med Res.
2019; 12(1): 6–11.
28.
Gunasekaran D, Thada R, Jeyakumar GFS, Manimegalai NP,
Shanmugam G, Sivagnanam UT. Physicochemical characterization
and self-assembly of human amniotic membrane and umbilical cord
collagen: A comparative study. Int J Biol Macromol. 2020; 165:
2920–33.
29.
Anusavice KJ, Shen C, Rawls HR. Phillips’ science of dental
materials. 12th ed. Philadelpia: Saunders; 2013. p. 592.
30.
Ari MDA, Yuliati A, Rahayu RP, Saraswati D. The differences
scaffold composition in pore size and hydrophobicity properties
as bone regeneration biomaterial. J Int Dent Med Res. 2018; 11(1):
318–22.
31.
Lee J, Yun H. Hydroxyapatite-containing gelatin/chitosan
microspheres for controlled release of lysozyme and enhanced
cytocompatibility. J Mater Chem B. 2014; 2(9): 1255–63.
32.
Wang Q, Wang Q, Wan C. The effect of porosity on the structure and
properties of calcium polyphosphate bioceramics. Ceram - Silikaty.
2011; 55(1): 43–8.
Copyright © 2023 Dental Journal (Majalah Kedokteran Gigi) p-ISSN: 1978-3728; e-ISSN: 2442-9740. Accredited No. 158/E/KPT/2021.
Open access under CC-BY-SA license. Available at https://e-journal.unair.ac.id/MKG/index
DOI: 10.20473/j.djmkg.v56.i3.p172–177