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Value of human amniotic epithelial cells in tissue engineering for cornea

Fatimah SS, Ng SL, Chua KH, Hayati AR, Tan AE, Tan GC

Hum. Cell, 2010 Nov;23(4):141-51.
PMID: 21166885 DOI: 10.1111/j.1749-0774.2010.00096.x

Human amniotic epithelial cells (hAECs) are potentially one of the key players in tissue engineering due to their easy availability. The aim of the present study was to develop an optimal isolation and transportation technique, as well as to determine the immunophenotype and epithelial gene expression of hAECs. Amnion was mechanically peeled off from the chorion and digested with trypsin-ethylenediaminetetraacetic acid. The isolated hAECs were cultured in medium containing 10 ng/mL epidermal growth factor until P4. The epithelial gene expression, cell surface antigen and protein expression of hAECs were analyzed by quantitative polymerase chain reaction, flow cytometry and immunocytochemistry. hAECs were also cultured in adipogenic, osteogenic and neurogenic induction media. The best cell yield of hAECs was seen in the digestion of 15 pieces of amnion (2 × 2 cm) and isolated 30 min after digestion with trypsin. F12:Dulbecco's modified eagle medium was the best medium for short term storage at 4 °C. hAECs expressed CD9, CD44, CD73 and CD90, and negligibly expressed CD31, CD34, CD45 and CD117. After serial passage, CK3, CK19 and involucrin gene expressions were upregulated, while p63, CK1 and CK14 gene expressions were downregulated. Sustained gene expressions of integrin β1 and CK18 were observed. At initial culture, these cells might have stem-like properties. However, they differentiated after serial passage. Nonetheless, hAECs have epithelial stem cell characteristics and have the potential to differentiate into corneal epithelial cells. Further investigations are still needed to elucidate the mechanism of differentiation involved and to optimize the culture condition for long term in vitro culture.

Matched MeSH terms: Amnion/cytology*
Fulltext Underlay myringoplasty: comparison of human amniotic membrane to temporalis fascia graft

Harvinder S, Hassan S, Sidek DS, Hamzah M, Samsudin AR, Philip R

Med J Malaysia, 2005 Dec;60(5):585-9.
PMID: 16515109

Human amniotic membrane as a homograft material was compared to temporalis fascia to close tympanic membrane perforations in 50 patients with chronic otitis media. Human amniotic membrane was used in 20 patients while temporalis fascia was used in the remaining 30. Anatomical closure of the perforation and reduction of the air-bone gap was measured. The graft uptake showed a 65% success rate for the amniotic membrane and 56.7% for the temporalis fascia at 3 months post-operatively. Significant closure of air-bone gap was observed in the human amniotic group. These results indicate comparable outcomes between human amniotic membrane and the temporalis fascia graft.

Matched MeSH terms: Amnion/transplantation*
Fulltext Therapeutic effects of conditioned - DPBS from amniotic stem cells on lactating cow mastitis

Ting WJ, Shaw SW, Hii LY, Lin TY, Chang SC, Liu KY, et al.

Taiwan J Obstet Gynecol, 2020 Jul;59(4):520-526.
PMID: 32653123 DOI: 10.1016/j.tjog.2020.05.009

OBJECTIVE: Bovine mastitis results in economic loss due to decrease in milk production. Antibiotic ointments are commonly used for treating. However, residue and anti-microbial resistance warranted attention progressively. Fortunately, stem cell anti-inflammatory properties and paracrine expression of cytokines accelerates wound healing and suppresses inflammatory reactions in mastitis. The objective of this study is to use the conditioned-Dulbecco's pluripotent stem cells (DPBS) from amniotic membrane stem cells (AMSCs) in treating bovine mastitis.
MATERIALS AND METHODS: The cows with mastitis were divided into two groups. In antibiotic control group, the cows were given tetraneomycin ointment. In conditioned-DPBS of AMSCs treatment group, amniotic membrane was collected for AMSCs after delivery. With expression of surface antigen and potential of tri-linage differentiation, AMSCs were injected into mammary glands. Then, milk was sampled every three days to monitor the effect of both treatments. The quality of milk was measured with pH, titratable acidity, free calcium ions and somatic cell count.
RESULTS: Our results demonstrated the Bovine AMSCs expressed CD44, low levels of CD4 and no CD105. Bovine AMSCs demonstrated the differentiation capability in the tri-cell lineages. Mastitis treatment with conditioned-DPBS from AMSCs (experimental group) and conventional antibiotics (control group) showed insignificant difference in pH value and titratable acidity. The level of ionic calcium concentration in the conditioned-DPBS group decreased from 3rd day to 12th day, while the level in the antibiotic group decreased from 0 day to 12th day. The somatic cell number was similar in both groups, which meet the standard of Taiwan milk collection.
CONCLUSION: In conclusion, conditioned-DPBS from bovine AMSCs has the therapeutic potential to treat bovine mastitis and may replace antibiotics therapy in the future.

Matched MeSH terms: Amnion
The use of autologous fibrin as a scaffold for cultivating autologous conjunctiva in the treatment of conjunctival defect

Safinaz MK, Norzana AG, Hairul Nizam MH, Ropilah AR, Faridah HA, Chua KH, et al.

Cell Tissue Bank, 2014 Dec;15(4):619-26.
PMID: 24633432 DOI: 10.1007/s10561-014-9436-y

The purpose of this study was to compare the use of autologous fibrin to human amniotic membrane (HAM) as a scaffold in cultivating autologous conjunctiva for transplantation in treatment of conjunctival defect. An experimental study was performed using 18 adult New Zealand white strain rabbits which were divided into 3 groups. Each group consists of 6 rabbits. The conjunctiva on the temporal site was excised to create a conjunctival epithelial defect. The excised area in the Group 1 was transplanted with autologous conjunctiva cultivated on autologous fibrin; Group 2 was transplanted with autologous conjunctiva cultivated on HAM and Group 3 was left bare. The rabbits were followed up at regular intervals until 6 weeks. The mean period of complete conjunctival epithelization was 11.50 ± 8.22 days for the autologous fibrin group, 15.33 ± 11.80 days for the HAM group and 25.33 ± 5.32 days in the bare sclera group. The epithelization rate for the autologous fibrin group was faster compared to the other two groups. However all the results were not statistically significant (p value >0.05). There were no postoperative complications noted during the follow up. Autologous fibrin is comparable to HAM as a scaffold for cultivation of conjunctiva in the treatment of conjunctival defect.

Matched MeSH terms: Amnion
The surgical induction of labour

LLEWELLYN-JONES D

Med J Malaya, 1958 Sep;13(1):80-5.
PMID: 13589375
Matched MeSH terms: Amnion*
The stemness gene expression of cultured human amniotic epithelial cells in serial passages

Simat SF, Chua KH, Abdul Rahman H, Tan AE, Tan GC

Med J Malaysia, 2008 Jul;63 Suppl A:53-4.
PMID: 19024980

The aim of the study is to evaluate the stemness gene expression of cultured human amniotic epithelial cells (HAECs) in serial passages. HAECs obtained from human term placentae were cultured in F12:DMEM(1:1) + 10% FBS +10ng/ml EGF in serial passages (P0, P1, P2 and P4). Quantitative RT-PCR was used to assess the gene expression analysis. The results showed that cultured HAECs expressed and downregulated the stemness genes expression for Oct-4, Sox-2, Nanog3, FGF4, Rex-1, FZD-9, BST-1 ABCG2. However, vimentin and nestin gene expression were upregulated. The results suggested that cultured HAECs may have pluripotent and multipotent properties.

Matched MeSH terms: Amnion/cytology*
Stemness and angiogenic gene expression changes of serial-passage human amnion mesenchymal cells

Fatimah SS, Tan GC, Chua K, Fariha MM, Tan AE, Hayati AR

Microvasc Res, 2013 Mar;86:21-9.
PMID: 23261754 DOI: 10.1016/j.mvr.2012.12.004

Particular attention has been directed towards human amnion mesenchymal stem cells (HAMCs) due to their accessibility, availability and immunomodulatory properties. Therefore, the aim of the present study was to determine the temporal changes of stemness and angiogenic gene expressions of serial-passage HAMCs.

Matched MeSH terms: Amnion/cytology*
Serial membrane sweeping at term in planned vaginal birth after cesarean: a randomized controlled trial

Hamdan M, Sidhu K, Sabir N, Omar SZ, Tan PC

Obstet Gynecol, 2009 Oct;114(4):745-751.
PMID: 19888030 DOI: 10.1097/AOG.0b013e3181b8fa00

OBJECTIVE: To estimate the effect of serial membrane sweeping on the onset of labor in women who planned vaginal birth after cesarean (VBAC).
METHODS: Women at term with one transverse lower segment cesarean delivery who were suitable for and who planned VBAC were approached to participate. Participants were randomly assigned to weekly membrane sweeping or weekly vaginal assessment for Bishop score until delivery. Participants and delivery providers were blinded to the allocated treatment. Standard obstetric care was given to all participants. The primary outcome was onset of labor which was defined as the presence of spontaneous regular and painful contractions that cause cervical dilation to at least 3 cm or prelabor rupture of membranes. Secondary outcomes included induction of labor and repeat cesarean delivery.
RESULTS: One hundred eight women were randomly assigned to membrane sweeping and 105 to control. The spontaneous labor rate was 78.5% compared with 72.1% (relative risk [RR] 1.1, 95% confidence interval [CI] 0.9-1.3; P=.34), the induction of labor rate was 12.1% compared with 9.6% (RR 1.3, 95% CI 0.6-2.8; P=.66), and the all-cause cesarean delivery rate was 40.2% compared with 44.2% (RR 0.9, 95% CI 0.7-1.2; P=.58) for the membrane sweeping and control groups, respectively. Gestational age at delivery (mean+/-standard deviation) of 39.6+/-1.0 weeks for the membrane sweeping group compared with 39.6+/-0.9 weeks for the control group (P=.84) was no different.
CONCLUSION: Serial membrane sweeping at term in women who planned VBAC has no significant effect on the onset of labor, pregnancy duration, induction of labor, or repeat cesarean delivery.
CLINICAL TRIAL REGISTRATION: ISRCTN, isrctn.org, ISRCTN55163179.
LEVEL OF EVIDENCE: I.

Matched MeSH terms: Amnion*
Scanning electron microscopic assessment on surface morphology of preserved human amniotic membrane after gamma sterilisation

Ab Hamid SS, Zahari NK, Yusof N, Hassan A

Cell Tissue Bank, 2014 Mar;15(1):15-24.
PMID: 23187886 DOI: 10.1007/s10561-012-9353-x

Human amniotic membrane that has been processed and sterilised by gamma irradiation is widely used as a biological dressing in surgical applications. The morphological structure of human amniotic membrane was studied under scanning electron microscopy (SEM) to assess effects of gamma radiation on human amniotic membrane following different preservation methods. The amniotic membrane was preserved by either air drying or submerged in glycerol before gamma irradiated at 15, 25 and 35 kGy. Fresh human amniotic membrane, neither preserved nor irradiated was used as the control. The surface morphology of glycerol preserved amnion was found comparable to the fresh amniotic membrane. The cells of the glycerol preserved was beautifully arranged, homogonous in size and tended to round up. The cell structure in the air dried preserved amnion seemed to be flattened and dehydrated. The effects of dehydration on intercellular channels and the microvilli on the cell surface were clearly seen at higher magnifications (10,000×). SEM revealed that the changes of the cell morphology of the glycerol preserved amnion were visible at 35 kGy while the air dried already changed at 25 kGy. Glycerol preservation method is recommended for human amniotic membrane as the cell morphological structure is maintained and radiation doses lower than 25 kGy for sterilization did not affect the appearance of the preserved amnion.

Matched MeSH terms: Amnion/radiation effects*; Amnion/transplantation*
Recurrence of peripheral ulcerative keratitis on the corneoscleral button in a young man treated successfully with oral corticosteroids

Wahid WA, Selvaraja V, Bakiah S, Ibrahim M

Cornea, 2008 Aug;27(7):837-9.
PMID: 18650673 DOI: 10.1097/ICO.0b013e318169d6cc

To describe recurrent peripheral ulcerative keratitis (PUK) on the corneoscleral graft in a young man treated successfully with oral corticosteroids.

Matched MeSH terms: Amnion/transplantation
Reconstruction of limbal stem cell deficient corneal surface with induced human bone marrow mesenchymal stem cells on amniotic membrane

Rohaina CM, Then KY, Ng AM, Wan Abdul Halim WH, Zahidin AZ, Saim A, et al.

Transl Res, 2014 Mar;163(3):200-10.
PMID: 24286920 DOI: 10.1016/j.trsl.2013.11.004

The cornea can be damaged by a variety of clinical disorders or chemical, mechanical, and thermal injuries. The objectives of this study were to induce bone marrow mesenchymal stem cells (BMSCs) to corneal lineage, to form a tissue engineered corneal substitute (TEC) using BMSCs, and to treat corneal surface defects in a limbal stem cell deficiency model. BMSCs were induced to corneal lineage using limbal medium for 10 days. Induced BMSCs demonstrated upregulation of corneal stem cell markers; β1-integrin, C/EBPδ, ABCG2, and p63, increased protein expression of CK3 and p63 significantly compared with the uninduced ones. For TEC formation, passage 1 BMSCs were trypsinized and seeded on amniotic membrane in a transwell co-culture system and were grown in limbal medium. Limbal stem cell deficiency models were induced by alkaline injury, and the TEC was implanted for 8 weeks. Serial slit lamp evaluation revealed remarkable improvement in corneal regeneration in terms of corneal clarity and reduced vascularization. Histologic and optical coherence tomography analyses demonstrated comparable corneal thickness and achieved stratified epithelium with a compact stromal layer resembling that of normal cornea. CK3 and p63 were expressed in the newly regenerated cornea. In conclusion, BMSCs can be induced into corneal epithelial lineage, and these cells are viable for the formation of TEC, to be used for the reconstruction of the corneal surface in the limbal stem cell deficient model.

Matched MeSH terms: Amnion/cytology*
Raised prorenin and renin concentrations in pre-eclamptic placentae when measured after acid activation

Singh HJ, Rahman A, Larmie ET, Nila A

Placenta, 2004 Aug;25(7):631-6.
PMID: 15193869

The aim of the study was to ascertain if there was any difference in the levels of prorenin and active renin between pre-eclamptic and normotensive feto-placental tissues.

Matched MeSH terms: Amnion/chemistry
Quantitative RT PCR approach to evaluate the neurogenic and gliagenic gene expression of cultured human amniotic epithelial cells

Tan GC, Simat SF, Abdul Rahman H, Tan AE, Chua KH

Med J Malaysia, 2008 Jul;63 Suppl A:51-2.
PMID: 19024979

The aim of the study is to determine the neuronal and glial gene expression of cultured human amniotic epithelial cells (HAECs) in serial passages. HAECs obtained from human term placentae were cultured in F12:DMEM (1:1) + 10% FBS +10ng/ml EGF in serial passages. Quantitative RT-PCR was used to assess the gene expression analysis. The results showed that the cultured HAECs expressed the neural stem cell genes (Nestin, NSE and Vimentin), mature neuronal genes (TH, MAP-2, beta-III-tubulin and NFM) and glial genes (CNPase, MBP and Olig). These neural stem cell genes increased with serial passages while the genes expression for mature neuronal and glial cells were downregulated. These results suggested that HAECs may promote or involve in neurogenesis and gliagenesis.

Matched MeSH terms: Amnion/cytology*
Quantifying the ultrastructure changes of air-dried and irradiated human amniotic membrane using atomic force microscopy: a preliminary study

Mohd S, Ghazali MI, Yusof N, Sulaiman S, Ramalingam S, Kamarul T, et al.

Cell Tissue Bank, 2018 Dec;19(4):613-622.
PMID: 30056604 DOI: 10.1007/s10561-018-9711-4

Air-dried and sterilized amnion has been widely used as a dressing to treat burn and partial thickness wounds. Sterilisation at the standard dose of 25 kGy was reported to cause changes in the morphological structure as observed under the scanning electron microscope. This study aimed to quantify the changes in the ultrastructure of the air-dried amnion after gamma-irradiated at several doses by using atomic force microscope. Human placentae were retrieved from mothers who had undergone cesarean elective surgery. Amnion separated from chorion was processed and air-dried for 16 h. It was cut into 10 × 10 mm, individually packed and exposed to gamma irradiation at 5, 15, 25 and 35 kGy. Changes in the ultrastructural images of the amnion were quantified in term of diameter of the epithelial cells, size of the intercellular gap and membrane surface roughness. The longest diameter of the amnion cells reduced significantly after radiation (p amnion cells reduced in the same direction without affecting the gross ultrastructure. At 15 kGy the intercellular gap decreased significantly (p

Matched MeSH terms: Amnion/radiation effects*; Amnion/ultrastructure*
Fulltext Outcome of Various Pterygium Excision Techniques in Tertiary Ophthalmic Centre in East Coast Malaysia

Khairidzan, M.K., Fatimah, S.S., Thangasamy, V.K.

International Medical Journal Malaysia, 2006;5(1):1-2.
MyJurnal

Pterygium is a common external eye problem. It is more frequently seen in tropical areas regions where exposure to ultraviolet sunlight is high. Clinically, a pterygium is a wing shaped fibrovascular growth arising from the bulbar conjunctiva onto the superficial cornea. Complications of pterygium include decreased in visual acuity, dryness, foreign body sensation and persistent redness. Surgical management is the mainstay of treatment for this condition. Numerous surgical techniques have been described in the treatment of pterygium. They include the bare sclera technique, simple direct conjunctival closure, rotational conjunctival graft and conjunctival autograft. Additional treatment to some of these techniques may include the use of beta particle therapy and antimetabolite therapy. Despite the wide range of surgical procedures described for its treatment, the main concern from these procedures has been the recurrence, which could be as high as 30% to 70%. Recurrent pterygium is often accompanied by increased conjunctival inflammation and accelerated corneal involvement. Repeated surgical procedures often only worsen the situation, as loss of conjunctival tissue and scarring can result in obliteration of the fornices and mechanical restriction of extraocular movements, with clinically significant diplopia. In Hospital Tengku Ampuan Afzan, pterygium excision is the most common surgical procedure after cataract extraction. We reviewed patients who had undergone pterygium surgery in HTAA in order to determine the most effective surgical method that could minimize recurrence. PURPOSE: To compare success rates of various excision techniques performed for primary and recurrent pterygium in Hospital Tengku Ampuan Afzan, Kuantan, Pahang.
METHODS: The outcome of 47 cases of pterygia (44 primary and 3 recurrent) excised with various techniques between January 2004 to September 2004 was retrospectively reviewed. Six clinical specialists and four trainees performed the surgical procedures. Outcome was evaluated in terms of recurrence of pterygia onto the cornea. RESULTS: The mean follow up was 3.04 months (range, 1-7 months). All pterygia were morphologically graded as intermediate or fleshy type except one. Four types of pterygium excision techniques were performed. Twenty-four cases had bare sclera, seventeen cases had conjunctival autograft transplantation, six cases had direct conjunctival closure and one had amniotic membrane transplantation done. Recurrence of pterygia occurred in thirteen eyes. Twelve cases from primary pterygia group and one case from recurrent group recurred. Recurrence rate was noted to be higher in direct conjunctival closure (4 out of 6 cases) and lowest in conjunctival autograft transplantation (2 out of 17 cases). Recurrence rate for bare sclera technique was noted to rank second in this study (6 out of 24 cases). In five cases of recurrence, subconjunctival tissue invasions were more than 1 mm but further surgical interventions were not needed at the time of this review was done. CONCLUSIONS: Conjunctival autografting was found to have less recurrent rate when compared with other techniques. The bare sclera technique was quoted to be associated with higher recurrence rate in other literatures. Interestingly in our series, recurrence rate for direct conjunctival closure technique was higher when compared to the former technique. This may be related to inadequate excision of pterygia tissue, which led to direct apposition of abnormal tissue to the cornea in the direct conjunctival closure technique. Even though the bare sclera technique is associated with a higher recurrence rate, it is still the preferred excision technique. This could be because it is less time consuming and technically easier to perform. Based on this study, conjunctival autografting should be the surgical procedure of choice for pteryigum in order to minimise the risk of recurrence.

Matched MeSH terms: Amnion
Organotypic culture of human amnion cells in air-liquid interface as a potential substitute for skin regeneration

Fatimah SS, Chua K, Tan GC, Azmi TI, Tan AE, Abdul Rahman H

Cytotherapy, 2013 Aug;15(8):1030-41.
PMID: 23830235 DOI: 10.1016/j.jcyt.2013.05.003

The aim of the present study was to evaluate the effects of air-liquid interface on the differentiation potential of human amnion epithelial cells (HAECs) to skin-like substitute in organotypic culture.

Matched MeSH terms: Amnion/cytology*
Oral prostaglandin e2 and amniotomy for induction of labour

Adeeb N, Fong TN

Med J Malaysia, 1974 Jun;28(4):263-6.
PMID: 4278401
Matched MeSH terms: Amnion/surgery
Immediate compared with delayed oxytocin after amniotomy labor induction in parous women: a randomized controlled trial

Tan PC, Soe MZ, Sulaiman S, Omar SZ

Obstet Gynecol, 2013 Feb;121(2 Pt 1):253-259.
PMID: 23344273 DOI: 10.1097/AOG.0b013e31827e7fd9

OBJECTIVE: To compare immediate with delayed (4 hours) oxytocin infusion after amniotomy on vaginal delivery within 12 hours and patient satisfaction with the birth process.
METHODS: Parous women with favorable cervixes after amniotomy for labor induction were randomized to immediate titrated oxytocin or placebo intravenous infusion in a double-blind noninferiority trial. After 4 hours, study infusions were stopped, the women were assessed, and open-label oxytocin was started if required. Maternal satisfaction with the birth process was assessed with a 10-point visual numerical rating scale (lower score, greater satisfaction).
RESULTS: Vaginal delivery rates at 12 hours were 91 of 96 (94.8%) compared with 91 of 94 (96.8%) (relative risk 0.98, 95% confidence interval [CI] 0.92-1.04, P=.72), and maternal satisfaction on a visual numerical rating scale (median [interquartile range]) was 3 [3-4] compared with 3 [3-5], P=.36 for immediate compared with delayed arm, respectively). Cesarean delivery, maternal fever, postpartum hemorrhage, uterine hyperactivity, and adverse neonatal outcome rates were similar between arms. The immediate oxytocin arm had a shorter amniotomy-to-delivery interval of 5.3±3.1 compared with 6.9±2.9 hours (P

Matched MeSH terms: Amnion/surgery*
Human amniotic membrane as a chondrocyte carrier vehicle/substrate: in vitro study

Krishnamurithy G, Shilpa PN, Ahmad RE, Sulaiman S, Ng CL, Kamarul T

J Biomed Mater Res A, 2011 Dec 01;99(3):500-6.
PMID: 21913317 DOI: 10.1002/jbm.a.33184

Human amniotic membrane (HAM) is an established biomaterial used in many clinical applications. However, its use for tissue engineering purposes has not been fully realized. A study was therefore conducted to evaluate the feasibility of using HAM as a chondrocyte substrate/carrier. HAMs were obtained from fresh human placenta and were process to produced air dried HAM (AdHAM) and freeze dried HAM (FdHAM). Rabbit chondrocytes were isolated and expanded in vitro and seeded onto these preparations. Cell proliferation, GAG expression and GAG/cell expression were measured at days 3, 6, 9, 12, 15, 21, and 28. These were compared to chondrocytes seeded onto plastic surfaces. Histological analysis and scanning electron microscopy was performed to observe cell attachment. There was significantly higher cell proliferation rates observed between AdHAM (13-51%, P=0.001) or FdHAM (18-48%, p = 0.001) to chondrocytes in monolayer. Similarly, GAG and GAG/cell expressed in AdHAM (33-82%, p = 0.001; 22-60%, p = 0.001) or FdHAM (41-81%, p = 0.001: 28-60%, p = 0.001) were significantly higher than monolayer cultures. However, no significant differences were observed in the proliferation rates (p = 0.576), GAG expression (p = 0.476) and GAG/cell expression (p = 0.135) between AdHAM and FdHAM. The histology and scanning electron microscopy assessments demonstrates good chondrocyte attachments on both HAMs. In conclusion, both AdHAM and FdHAM provide superior chondrocyte proliferation, GAG expression, and attachment than monolayer cultures making it a potential substrate/carrier for cell based cartilage therapy and transplantation.

Matched MeSH terms: Amnion/metabolism*; Amnion/ultrastructure
Human amnion as a novel cell delivery vehicle for chondrogenic mesenchymal stem cells

Tan SL, Sulaiman S, Pingguan-Murphy B, Selvaratnam L, Tai CC, Kamarul T

Cell Tissue Bank, 2011 Feb;12(1):59-70.
PMID: 19953328 DOI: 10.1007/s10561-009-9164-x

This study investigates the feasibility of processed human amnion (HAM) as a substrate for chondrogenic differentiation of mesenchymal stem cells (MSCs). HAM preparations processed by air drying (AD) and freeze drying (FD) underwent histological examination and MSC seeding in chondrogenic medium for 15 days. Monolayer cultures were used as control for chondrogenic differentiation and HAMs without cell seeding were used as negative control. Qualitative observations were made using scanning electron microscopy analysis and quantitative analyses were based on the sulfated glycosaminoglycans (GAG) assays performed on day 1 and day 15. Histological examination of HAM substrates before seeding revealed a smooth surface in AD substrates, while the FD substrates exhibited a porous surface. Cell attachment to AD and FD substrates on day 15 was qualitatively comparable. GAG were significantly highly expressed in cells seeded on FD HAM substrates. This study indicates that processed HAM is a potentially valuable material as a cell-carrier for MSC differentiation.

Matched MeSH terms: Amnion/cytology; Amnion/metabolism*; Amnion/ultrastructure

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