Stem Cell Injection into the Meibomian Glands to Restore Meibomian Gland Function

Stem Cell Injection into the Meibomian Glands to Restore Meibomian Gland Function

We are preparing to submit an IRB for this research project: injecting a patient's own stem cells into his or her own meibomian glands. My previous post has also more information: http://eyedoc2020.blogspot.com/2017/01/how-to-regenerate-stem-cells.html


The key questions are the following:
1. Does autologous serum contain enough stem cells to regenerate Meibomian Glands or is Adipose Fat or Bone Marrow stem cells better?
2. What are the risks of injecting stem cells in the Meibomian Glands?
3. How exactly will this be done?

My Hypothesis to these questions are as follows:
1. Does autologous serum contain enough stem cells to regenerate Meibomian Glands or is Bone Marrow stem cells better?

Adipose fat appears to have more concentrated stem cells than Bone Marrow. See this link and information below: http://stemcellrevolution.com/about-us/faqs/#12
I have not seen a paper yet to prove this: still looking:

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Whether your adult mesenchymal stem cells come from bone marrow or from fat probably does not make a difference in terms of clinical results. Although some centers claim that bone marrow derived cells are superior to fat derived cells, there is no evidence to substantiate that. Recent studies show that fat derived cells make bone tissue much better than the bone marrow derived cells. Some studies are showing different outcomes but it is important to realize that these studies are all done in petri dishes and may not relate to living organism. Also, it is important that one is not mislead in some marketing materials by the word “bone” in “bone marrow”, possibly implying that since this is an “orthopedic source” it must be better for treating orthopedic conditions such as cartilage regeneration. In fact, the bone marrow is part of the reticulo-endothelial system (makes blood cells) and just happens to be found in the center of bone. The truth is, both bone marrow derived and stromal (from fat) derived stem cells are both effective for regenerative therapy and both have the potential to differentiate into mature functional cartilage. However, stem cells from fat are 100 to 1000 times more plentiful and this makes same day procedures (allowed in the US) much more effective with fat derived cells. The higher numbers of cells in fat leads to better clinical outcomes. Also, the quality of bone marrow declines with age and it has less numbers of cells and less healthy cells compared to the fat. The diminution in quantity and quality of bone marrow cells related to age and chronic illness is well documented. Last but not least, the ease of removing fat from under the skin using a mini-liposuction under local anesthetic is much less invasive and MUCH LESS painful than undergoing bone marrow aspiration to obtain bone marrow cells.

How is the fat obtained?

CSN patients have their fat (usually abdominal) harvested in our special sterile harvesting technology under a local anesthetic. The “mini liposuction” fat removal procedure lasts approximately twenty minutes. Specially designed equipment is used to harvest the fat cells and less than 100cc of fat is required. Post-operative discomfort is minimal and there is minimal restriction on activity. RETURN TO TOP…

How does CSN control sterility in the processing of the stem cells?

Stem cells are harvested under sterile conditions using a special “closed system” technology so that the cells never come into contact with the environment throughout the entire process from removal to deployment. Sterile technique and oral antibiotics are also used to prevent infection. RETURN TO TOP…

Does CSN use stem cells from any other animal source or person?

No. Only a person’s own adult autologous cells are used. These are harvested from each individual and deployed back into their own body. There is no risk of contamination or risk of introduction of mammalian DNA into the treatments. RETURN TO TOP…

Why do some stem cell facilities around the world require days to weeks before removal of mesenchymal stem cells until reinsertion into the patient?

These facilities are obtaining stem cells from bone marrow or blood in relatively small quantities and they are then culturing (growing) the cells to create adequate quantities. Research seems to indicate that success of treatment is directly related to the quantity of cells injected. CSN uses adipose derived stem cells that are abundant naturally at approximately 2,500 times levels found in bone marrow (the most common source of mesenchymal stem cells). CSN uses technology that isolates adipose stem cells in vast numbers in a short time span so that prolonged culturing is unnecessary and cells can be deployed into a patient within 90 minutes of harvesting. 

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The IRB will have a protocol to inject one eye lid with Autologous Serum derived stem cells and another lid to use Bone Marrow Stem cells.
2. What are the risks of injecting stem cells in the Meibomian Glands?

Based on the research previously performed injecting stem cells into the spinal cord and brain area for patients with cerebral palsy and many other brain injury issues, I suspect there is little risk to the Meibomian glands or eye. I suspect the risk to the eye is close to zero. The risk of infection is the only risk I suspect but I have not seen yet a case of infection when stem cells were injected into the heart or brain areas.

3. How exactly will this be done?
The protocol will include preparing Autologous Serum and Bone Marrow Derived Stem Cells.
Probing and expression will be done first and then injection into as many meibomian gland orifices as possible after cleaning area with Betadine and giving anesthesia as we currently do.

More soon.

Sandra Lora Cremers, MD, FACS

Stem Cells Int. 2017;2017:3134543. doi: 10.1155/2017/3134543. Epub 2017 Mar 2.

Delivery of Bone Marrow-Derived Mesenchymal Stem Cells Improves Tear Production in a Mouse Model of Sjögren's Syndrome.

Aluri HS1, Samizadeh M1, Edman MC2, Hawley DR1, Armaos HL1, Janga SR2, Meng Z3, Sendra VG4, Hamrah P5, Kublin CL1, Hamm-Alvarez SF6, Zoukhri D7.

Author information

Abstract

The purpose of the present study was to test the potential of mouse bone marrow-derived mesenchymal stem cells (BD-MSCs) in improving tear production in a mouse model of Sjögren's syndrome dry eye and to investigate the underlying mechanisms involved. NOD mice (n = 20) were randomized to receive i.p. injection of sterile phosphate buffered saline (PBS, control) or murine BD-MSCs (1 × 106 cells). Tears production was measured at baseline and once a week after treatment using phenol red impregnated threads. Cathepsin S activity in the tears was measured at the end of treatment. After 4 weeks, animals were sacrificed and the lacrimal glands were excised and processed for histopathology, immunohistochemistry, and RNA analysis. Following BD-MSC injection, tears production increased over time when compared to both baseline and PBS injected mice. Although the number of lymphocytic foci in the lacrimal glands of treated animals did not change, the size of the foci decreased by 40.5% when compared to control animals. The mRNA level of the water channel aquaporin 5 was significantly increased following delivery of BD-MSCs. We conclude that treatment with BD-MSCs increases tear production in the NOD mouse model of Sjögren's syndrome. This is likely due to decreased inflammation and increased expression of aquaporin 5.

Cell Med. 2016 Oct 18;8(3):63-77. doi: 10.3727/215517916X693366. eCollection 2016.

Allogeneic Mesenchymal Stem Cell Transplantation in Dogs With Keratoconjunctivitis Sicca.

Bittencourt MK1, Barros MA2, Martins JF2, Vasconcellos JP1, Morais BP2, Pompeia C3, Bittencourt MD4, Evangelho KD5, Kerkis I3, Wenceslau CV3.

Author information

Abstract

Keratoconjunctivitis sicca (KCS) is a dysfunction in tear production associated with clinical signs, which include conjunctival hyperemia, ocular discharge, discomfort, pain, and, eventually, corneal vascularization and pigmentation. Immunosuppressive drugs are routinely administrated for long periods to treat KCS but with side effects and limited results. Evaluation of the clinical benefits of intralacrimal transplantation of allogeneic mesenchymal stem cells (MSCs) in dogs with mild-moderate and severe KCS was done. A total of 24 eyes with KCS from 15 dogs of different breeds were enrolled in the present study. A single transplantation of MSCs (1 × 106) directly into lacrimal glands (dorsal and third eyelid) was performed. The Schirmer tear tests (STTs) and ocular surface improvements were used to assess short- and long-term effects of these cells. The STTs were carried out on day 0 (before MSCs transplantation) and on days 7, 14, 21, and 28, as well as 6 and 12 months after MSC transplantation. Our data demonstrate that allogeneic MSC transplantation in KCS dogs is safe since no adverse effects were observed immediately after transplantation and in short- and long-term follow-ups. A statistically significant increase in the STT and ocular surface improvements was found in all eyes studied. In all the eyes with mild-moderate KCS, STT values reverted to those of healthy eyes, while in eyes with severe KCS, although complete reversion was not found, there was improvement in tear production and in other clinical signs. Our study shows that a single dose of a low number of MSCs can be used to treat KCS in dogs. In contrast to immunosuppressive drug use, MSC transplantation has an effect over a long period (up to 12 months), even after a single administration, and does not require daily drug administration.



Protocol:

Adipose Tissue-Derived MSCs

1. A total of three female 6- to 12-month-old healthy mongrel dogs were used to isolate adipose MSCs. Visceral (ovary fat) fat samples were collected during elective surgeries (surgery independent of the study). Before enrolment, dogs underwent routine clinical examination, hematologic evaluation (plasma proteins, red blood cells count, white blood cells count, platelet number and hemoglobin concentration), and viral screening.

2. After collection, fat fragments were transported in a cooler box, under strict control of temperature, in transport medium composed of Dulbecco’s modified Eagle’s medium high glucose (DMEM-H) and 500 U/ml streptomycin and 500 U/ml penicillin (Thermo Fisher Scientific Waltham, MA, USA)—the samples were processed within 2 h. Adipose tissue cells were isolated using a standard protocol based on fragmentation followed by collagenase IV digestion, following procedures described in Mambelli and coauthors27. The isolated cells were plated at 1 × 105 on 36-mm dishes (TPP, Trasadingen, Switzerland) with DMEM-H supplemented with 15% HyClone fetal bovine serum (Catalog No. SH30070-03; Logan, UT, USA), 100 U/ml streptomycin and 100 U/ml penicillin, 2 mM l-glutamine, and 1% nonessential amino acids (all Thermo Fisher Scientific), which is here designated as basal culture medium. The cultures were incubated at 37°C in a humidified atmosphere containing 5% CO2. After 4 to 7 days, cells were washed twice in phosphate-buffered saline (PBS; Gibco, Gaithersburg, MD, USA), dissociated with 0.25% trypsin (Thermo Fisher Scientific), and expanded in 75-cm2 culture flasks (TPP).