The Use of Autologous Serum in the Development of Corneal and Oral Epithelial Equivalents in Patients with Stevens-Johnson Syndrome

Nakamura, Takahiro; Ang, Leonard P. K.; Rigby, Helen; Sekiyama, Eiichi; Inatomi, Tsutomu; Sotozono, Chie; Fullwood, Nigel J.; Kinoshita, Shigeru

doi:10.1167/iovs.05-1188

Cited by 62 publications

(55 citation statements)

References 21 publications

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“…These cultured BMECs were negative for the corneal epithelium-specific marker K12 and hence do not acquire the corneal phenotype, confirming the earlier reports. 10,29 Corneal surface reconstruction Our study suggests that BMECT is a useful SC therapy for severe bilateral LSCD due to chemical injury as described earlier. 7,9,30 In addition, it is beneficial in some unilateral LSCD patients in whom either transplantation of autologous cultured limbal epithelium failed or was not possible (patients 3 and 4 in Table 3).…”

Section: Ex-vivo Expansion Of Bmescs In Culturesupporting

confidence: 65%

“…The present study confirms the earlier reports on the presence of SCs in the cultured buccal epithelium by their functional property of CFE and ability to form holoclones. 10,29 Moreover, we demonstrate for the first time an increase in the total number of SCs in the ex-vivo expanded buccal epithelium, although not the percentage of SCs. These cultured BMECs were negative for the corneal epithelium-specific marker K12 and hence do not acquire the corneal phenotype, confirming the earlier reports.…”

Section: Ex-vivo Expansion Of Bmescs In Culturementioning

confidence: 64%

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Adult human buccal epithelial stem cells: identification, ex-vivo expansion, and transplantation for corneal surface reconstruction

Priya

Arumugam

Vaishali

et al. 2011

Eye

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Purpose To identify adult human buccal epithelial stem cells (SCs) on the basis of two parameters (high p63 expression and greater nucleus/cytoplasmic (N/C) ratio) and to evaluate clinical efficacy of ex-vivo expanded autologous epithelium in bilateral limbal SC-deficient (LSCD) patients. Methods The epithelial cells were isolated from buccal biopsy and cultured on human amnion in culture inserts with 3T3 feeder layer. The SCs were identified on the basis of two-parameter analysis using confocal microscopy, surface markers, and colonyforming efficiency (CFE). The cultured epithelium was transplanted in 10 LSCD patients followed by penetrating keratoplasty in 4 patients. The clinical outcome was followed up to 3 years. Results A distinct population (3.0±1.7%) of small cells expressing high levels of p63 with greater N/C ratio was observed in buccal epithelium. The N/C ratio was found to be more appropriate than cell diameter for two-parameter analysis. These cells located in the basal layer were negative for connexin-43 and positive for melanoma-associated chondroitin sulfate proteoglycan, containing holoclones with 0.2% CFE, thus representing the SC population. After transplantation of cultured epithelium with increased (sixfold) SC content, anatomical and visual improvement was observed at 13-34 months in 3/10 LSCD patients. Conclusions The two-parameter SC marker is useful to identify and quantify buccal epithelial SCs. The transplantation of bioengineered SC-rich buccal epithelium is a strategy for corneal surface reconstruction in bilateral LSCD. However, further studies are required to optimize the culture conditions and to look for other sources of adult SCs for better visual outcome.

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Section: Ex-vivo Expansion Of Bmescs In Culturesupporting

confidence: 65%

Section: Ex-vivo Expansion Of Bmescs In Culturementioning

confidence: 64%

Adult human buccal epithelial stem cells: identification, ex-vivo expansion, and transplantation for corneal surface reconstruction

Priya

Arumugam

Vaishali

et al. 2011

Eye

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“…23,24 As the cultures were not air-lifted, epithelial stratification was less then reported earlier, 25,26 and the cells appeared generally flatter. Interestingly, transcription factor p63, 11 and notably low affinity NGF receptor p75, 27 the progenitor cell markers known to be expressed by oral mucosal epithelial cells were both found in the cultures, along with the expression of ABCG2.…”

Section: Discussionmentioning

confidence: 78%

Transplantation of cultivated oral mucosal epithelial cells for severe corneal burn

Kuo

et al. 2009

Eye

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Purpose To access the feasibility of using cultivated oral mucosal epithelial cell transplantation (COMET) for the management of severe corneal burn. Methods COMET was performed to promote re-epithelialization in two eyes with acute alkaline burn and one eye with chronic alkaline burn, and to reconstruct the ocular surface in two eyes with chronic thermal burn. Autologous oral mucosal epithelial cells obtained from biopsy were cultivated on amniotic membrane. Immunoconfocal microscopy for keratins and progenitor cell markers was performed to characterize the cultivated epithelial sheet. Following transplantation, the clinical outcome and possible complications were documented. The patients were followed for an averaged 29.6 ± 3.6 (range: 26-34) months. Results Cultivated oral mucosal epithelial sheet expressed keratin 3, 13, and progenitor cell markers p63, p75, and ABCG2. After COMET, all the corneas became less inflamed, and the corneal surface was completely reepithelialized in 6.0 ± 3.2 (range: 3-10) days in all but one patients. Microperforation occurred in one patient, and a small persistent epithelial defect developed in another. Both were solved uneventfully. In all patients, superficial corneal blood vessels invariably developed, and to further improve vision, conjunctivo-limbal autografting (N ¼ 3) and/or penetrating keratoplasty (N ¼ 3) were performed subsequently. The vision of all patients showed substantial improvement after additional surgeries. Conclusions This study showed the potential of COMET to promote re-epithelialization and reduce inflammation in acute corneal burn, and to reconstruct the corneal surface in chronic burn. COMET may, therefore, be considered an alternative treatment for severe corneal burn.

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“…Therefore, using oral keratinocytes as seed cells is the best choice. A great deal of research on culturing oral keratinocytes in vitro has been developed in the last 20 years [12][13][14] . However, there was always a lot of deficiencies or shortcomings.…”

Section: Color Version Available Onlinementioning

confidence: 99%

Preliminary Experimental Study of Tissue-Engineered Urethral Reconstruction Using Oral Keratinocytes Seeded on BAMG

Song

et al. 2008

Urol Int

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Introduction: This study investigates the feasibility of replacing urinary epithelial cells with oral keratinocytes seeded on a bladder acellular matrix graft (BAMG) to reconstruct a tissue-engineered urethra. Materials and Methods: After enzymatic treatment of a small segment of rabbit buccal mucosa (1.0 × 0.4 cm), the epidermis and dermis were separated mechanically. Oral keratinocytes were isolated from the epidermis and seeded onto a culture dish with a feeder layer of 3T3 mouse fibroblasts inhibited by mitomycin (i3T3) or on a culture dish without i3T3, respectively. These cells were expanded and applied to sterilized BAMG to obtain a tissue-engineered mucosa. Oral keratinocytes were assessed by growth curve, morphology and immunofluorescence staining. The tissue-engineered mucosa was assessed using morphology, immunohistochemistry staining and scanning electron microscopy. Results: Oral keratinocytes seeded onto a feeder layer of i3T3 had finer morphous and better amplification capability, and could be passaged for 8 generations. Oral keratinocytes that were cultured without i3T3 could only be subcultured 2 generations before ageing. Passage 2 oral keratinocytes cultured with i3T3 were expanded and seeded onto sterilized BAMG to obtain a tissue-engineered mucosa for urethral reconstruction. The compound graft was assessed using HE staining and scanning electron microscopy. Oral keratinocytes had good compatibility with BAMG. Conclusions: Rabbit oral keratinocytes of can be cultured in vitro and attain magnitude in quantity. Oral keratinocytes also had good compatibility with BAMG and the compound graft achieved could become a new choice in urethral reconstruction.

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The Use of Autologous Serum in the Development of Corneal and Oral Epithelial Equivalents in Patients with Stevens-Johnson Syndrome

Cited by 62 publications

References 21 publications

Adult human buccal epithelial stem cells: identification, ex-vivo expansion, and transplantation for corneal surface reconstruction

Adult human buccal epithelial stem cells: identification, ex-vivo expansion, and transplantation for corneal surface reconstruction

Transplantation of cultivated oral mucosal epithelial cells for severe corneal burn

Preliminary Experimental Study of Tissue-Engineered Urethral Reconstruction Using Oral Keratinocytes Seeded on BAMG

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