Morphological substantiation of application of cellular technologies for correction of striae

doi:10.4236/health.2010.212213

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Vol.2, No.12, 1430-1436 (2010) Health

doi:10.4236/health.2010.212213

Morphological substantiation of application of cellular

technologies for correction of striae

Igor Maiborodin*, Andrey Shevela, Andrey Babko, Vitaly Morozov, Vera Matveeva

Center of New Medical Technologies Institute of Chemical Biology and Fundamental Medicine, Novosibirsk, Russia;

*Corresponding Author: imai@mail.ru

Received 21 August 2010; revised 15 October 2010; accepted 19 October 2010

ABSTRACT

The possibility of the application of autologic

pluripotent cells from peripheral blood (APCPB)

for correction of striae after pregnancy was

studied. Visually first signs of improvement of

atroph ic scars can be noticed in 6-8 w eeks a fter

injection of APCPB and gradually progress

during 6 months. The atrophy of epithelium,

hyperkeratosis, sclerosis of all dermis layers

with smoothing of papillae, reduction of number

of blood vessels and skin appendages were

found before treatment by method of light mi-

croscopy. In all observations after injection of

APCPB in a connective tissue, which replaces

of dermis, an increase of number of vessels at

the expense of processes of neoangiogenesis,

leukocytic infiltration reduction, thinning and

ordering of arrangement of collagen and elastin

fibers were detected. These results give a ch a nc e

for more successful application of cosmetology

procedures for correction of these defects of

skin.

Keywords: Skin Striae after Pregnancy; Atrophy of

Epidermis; Dermal Sclerosis; Pluripotent Cells from

Peripheral Blood; Ne oangiogenesis

1. INTRODUCTION

Striae distensae or stretch marks—linear atrophic

scars, which are localized mainly on skin of mammary

glands, stomach, hips and buttocks. At the beginning of

the development they are bright pink in color, often with

a bluish shade. This color is due to active vascular inva-

sion—a period of fresh stretch marks, which lasts for

about 6-12 months. Then there is blanch ing of tissue, the

surface becomes shin y, in texture similar to filigree (cig-

arette paper). Striae color is lighter than surrounding

normal skin because scar tissue contains very few mela-

nocytes and melanin. For the same reason, when the

surrounding skin tans, white strips remain [1-3]. From

the medical point of view these scars are completely

harmless, but they are a widespread cosmetic problem.

The treatment of this disease has been a challenge for

clinicians and experimenters for a long time [4].

The pathogenesis of striae is still not completely

known, and possibly linked to changes in structures that

are responsible for elasticity a nd con tractility of the sk in.

These structures are the components of the extracellular

matrix, including fibrillin, elastin and various collagens.

In this process the important role plays decrease of ex-

pression of genes responsible for formation of collagen,

elastin and fibronectin, which, in turn, accompanied by

metabolic imbalance of fibroblasts, in particular

—metabolism of fibrillin in these cells [5-10]. It should

be emphasized that in striae were found signs of chronic

autoimmune disorders, in particular the reaction of “graft

versus host” [11].

In the time of light microscopy study of striae histo-

pathology were obtained data that well-formed elastin

fibers dominate in fresh defects, thickened fibers domi-

nate in the older parts of lesions. Most likely, the elastin

fibers in striae were synthesized again, they gradually

were thickened and their number increased. According to

the data of scanning electron microscopy, unusually

dense and well-developed net of elastin with horizontal

packing of collagen was discovered. Apparently, the

primary target of this pathological process is exactly

elastin fibers [12-14].

According to modern representations, physiological

regeneration of tissue in adult organism and its repara-

tion in case of damage happens with direct participation

of low differentiated cells-precursors or stem cells. The

basic source of stem cells is bone marrow, which in ad-

dition to the basic function—haemopoetic, is able to

generate precursors of cellular elements of a great num-

ber of tissues in organism.

Besides bone marrow the pluripotent cells were dis-

covered in other tissues of an adult organism—adiposal,

muscular and nerve tissue, and also in peripheral blood

I. Maiborodin et al. / Health 2 (2010) 1430-1436

1431

and umbilical cord/placen ta blood. Moreover, it is estab-

lished that depending on a microenvironment the stem

cells are able to get through haemopoetic/mesenchymal

barrier as possess to high plasticity concerning differen-

tiation and transdifferentiation.

2. AIM

Because of the small efficiency of widely applied

techniques of the treatment of striae has been made an

attempt to correct this pathology with use of cellular

technologies. We demonstrate the morphological de-

scription of several cases from practice.

3. MATERIAL AND METHODS

All actions, starting from material-source for receiv-

ing of pluripotent cells before the procedure of cellular

therapy, were made with observance of existing legal

and ethical standards of Russian Federation, and also

other standard documents and recommendations con-

cerning this area of medicine. The treatment of striae on

stomach skin after pregnancy was performed only after

receiving the informed written approval from patients. In

our work only autologic cells from peripheral blood

were used—as it is more practical and safer as does not

demand of selection of matching “donor-recipient”.

Isolation of a cellular material from autologic blood

received from an elbow vein was performed on the de-

vice for cytoplasmapheresis «Haemonetics» (MCS +, the

USA). This device uses a make-and-break current prin-

ciple through centrifuging chamber which was named

“bell”. There is a division of blood into components in

dependence on weight under the influence of centrifugal

force. Using this system the device is able to isolate any

cellular components from blood at donor or medical

apheresis.

Each component leaving a bell passes through an opti-

cal linear sensor which allows to define optical density of

medium. That makes possible in advance to program

separation of specific components of blood, such as plas-

ma, platelet s, lym phocy t es, granul ocy t es, st em cell s , et c.

The method of flow cytometry was used for the anal-

ysis of stem cell population containing in the received

material. Samples were analyzed on flow cytofluometer

«FACSAria» using FACSDiVa (Becton Dickinson)

software.

We used monoclonal antibodies specific to CD34,

conjugated with FITC; specific to CD38, conjugated

with Cy-Chrome (Becton Dickinson); to antigens to the

surface of human cells; isotypical controls to antibodies

of classes IgG1, IgG2a mice (Becton Dickinson), conju-

gated with FITC and PE, accordingly, under the instruc-

tion of the manufacturer.

According to the completed analysis on flow cytof-

luometer using antibodies to superficial human antigens

CD34 and CD38, the amount of CD34+ cells was 0.2%

and early precurso rs CD34+/CD38 was 0.1% out of total

population of cells. Besides, the quantity of mononuclear

cells in 1 ml was counted using trypan blue. We detected

that 1 ml of the sample, received after c yt op l as ma p h er e si s,

has contained 220 × 106 cells.

The received cellular material, containing autologic

pluripotent cells from peripheral blood (APCPB), was

entered directly after separation procedure into zones of

skin atrophy using following method:

1) Suspension of APCPB was collected into a sterile

syringe in capacity of 5.0 ml with the rubber pump. The

stroke of the syringe pump should be tight but smooth.

2) Before injection the skin was processed by a napkin

made of notwoven material wetted with 0.5% water so-

lution of chlorhexidine.

3) A needle on a syringe replaced with a needle 30G.

Injection was performed in zones of striae linearly retro-

gradually. A needle was entered on all length strictly by

cut upwards or cut downwards A needle was entered on

all length strictly by cut upward s or cut downwards. The

needle cut should not be placed sideways, that not to

injure the skin. During removal of the needle out of the

tissue the pressure upon the pump was made, for filling

skin atrophy by plasma containing autologic AHPCPB.

During procedure the needles were replaced several

times when they became blunt. Anesthesia is not neces-

sary, as the procedure is a little painful (diameter of the

needle is only 0.3 mm).

The samples of skin (epidermis and dermis) size 2 × 2

mm from the front surface of the stomach on border of

normal skin and striae after pregnancy were preserved in

a 4% paraformaldehyde on biphosphate buffer (pH 7.4)

for at least 24 hour, dehydrated in a gradien of ethanol,

lightened in xylene and embedded in histoplast. Micro-

scopic sections of 5-7 microns thick were stained with

hematoxylin and eosin, and studied under a light micro-

scope Axioimager M1 (Carl Zeiss, Germany) with a

magnification of up to 1200 times.

4. RESULTS

Patient O., 28 years old, (pregnancy was 4.5 years

ago), visually the first signs of improvement of atrophic

cicatrices appeared in 6-8 weeks after the injection of

AHPCPB. In 6 months filling striae from inside was well

visible (Figures 1-4). In the first place we noticed that

the color of striae approached the color of normal skin.

Thus “old” atrophic striae which did not acquire sun tan,

approached the color of normal skin and started to ac-

quire sun tan (Figure 2). The depth of tissue retraction

decreases. At this stage the changes of ski n were visible.

If initially skin was atrophic, thinned, at compression

easily gathers in folds and looks like wrinkled cigarette

paper (Figure 3), after 6 months it became more dense,

I. Maiborodin et al. / Health 2 (2010) 1430-1436

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Figure 1. Abdominal skin of the patient O. with numerous

linear atrophic scars before treatment.

Figure 2. Status of stretch marks in 6 months after the in-

troduction of APCPB. The color of the stretch mark is

much closer to the color of the surrounding skin, in some

areas there is pigmentation as a result of sun tan.

Figure 3. Abdominal skin with stretch marks after pregnancy

before using APCPB. Skin is atrophic, thinned, at compression

easily gathers in folds and looks like wrinkled cigarette paper.

Figure 4. 6 months after using AHPCPB the skin becomes

more dense, looks thicker, the striae become narrower.

looked thicker. Perhaps, because of the improvement of

surrounding skin the striae looked narrower (Figure 4).

In the time of histological study of striae of this pa-

tient, which was taken before the procedure of treatment

by APCPB, were noted an atrophy and thinning of epi-

thelium, hy perkerat osis, fl atteni ng of papil lae and smoothing

of papillary layer in dermis, and almost total absence of

reticular layer which was replaced with connective tissue

with a chaotic arrangement of fibers and a small amount

of blood vessels with perivascular leukocytic infiltration

with prevalence of lymphocytes and macrophages. The

type of vessels cannot always be identified due to expressed

sclerotic and inflammatory changes (Figures 5, 6).

In the study of biopsy from the edge of the defect

within 1 month after the injection of APCPB were also

found the above mentioned changes (atrophy of the epi-

thelium and hyperkeratosis, flattening and smoothing of

the dermal papillae, sclerosis of the reticular layer).

However, the severity of sclerotic changes in the dermis

was somewhat less, there was formation and order of

collagen fibers in the reticular layer, where was signifi-

cantly more blood vessels and drastically was reduced

degree of perivascular leukocyte infiltration. Also were

found young blood vessels with very thin walls, it was

not always possible to trace the endothelial line. Red

blood cells were present not only in these vessels, but

were also located around vessels (Figures 7, 8). These

newly formed blood vessels were not even remotely

similar to the granulations, an d therefore the assumption

of the induction of inflammation in tissues after APCPB

injection sh ould be rejected.

4 months after the injection of APCPB the atrophic

changes in the epidermis became more pronounced,

completely vanished skin appendages and smoothed

papillae, but one can note the appearance of melanocytes.

Signs of edema were more significant, there were cavi

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Figure 5. The results of morphological study of stretch marks

of the patient O. before treatment. Atrophy and thinning of the

epithelium, hyperkeratosis, flattening of the papillary layer in

dermis. Reticular layer is replaced by connective tissue with

randomly distributed fibers and a small amount of blood ves-

sels with perivascular leukocytic infiltration. Hematoxylin and

eosin. Magnification × 40.

Figure 6. The fragment of Figure 5. Chaotic arrangement of

fibers in the intercellular matrix of connective tissue in dermis.

In cytogram of perivascular leukocytic infiltrates the lympho-

cytes and macrophages are dominate. Magnification × 400.

ties with fluid, an expansion of the components of the

lymphatic vessels was found. Scar connective tissue in

the reticular layer of the dermis looked friable. The

number of vessels decreased slightly, but still remained

more than in initial conditions. The vessels became larg-

er and sclerosis of their walls can be noted. By that time

there were again appeared perivascular leukocytic infil-

tration with a predominance of lymphocytes. Young

vessels with thin walls were still present in the dermis

(Figur es 9, 10).

In skin biopsy of patient M., 22 years old (pregnancy

2.5 years ago), before treatment were found atrophy of

Figure 7. Biopsy from the edge of the defect within 1 month

after the injection of APCPB: atrophy of the epithelium,

hyperkeratosis, flattening of the dermal papillae, appendages of

the skin are absent. The severity of sclerotic transformation of

the reticular layer of the dermis is reduced, the fibers of inter-

cellular substance became thinner and arranged in parallel. The

number of vessels increased. Hematoxylin and eosin. Magnifi-

cation × 40.

Figure 8. The fragment of Figure 7. Drastic reduction of peri-

vascular leukocytic infiltration. In the connective tissue the

newly-formed blood vessels with thin walls are present , where

nuclei of endothelial cells are very rare. Red blood cells are

located not only in vessels but also perivascular. Magnification

× 400.

the epithelium, hyperkeratosis, and almost complete ab-

sence of all layers of the dermis—papillary and reticular,

and complete absence of skin appendages: hair follicles,

sudoriferous and sebaceous glands. The dermis was re-

placed with scar connective tissue with edema and a

small amount of blood vessels, around which the perivas-

cular leukocytic infiltration with a predominance of

lymphocytes and a large number of neutrophils was

present.

I. Maiborodin et al. / Health 2 (2010) 1430-1436

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In the study of biopsy specimen from the edge of the

Figure 9. Striae area in 4 months after injection APCPB.

Atrophic changes and hyperkeratosis of the epidermis are more

pronounced. Skin appendages vanished completely and papil-

lae are smoothed. Instead of scar connective tissue in the reti-

cular layer of the dermis there is friable connective tissue

where significant signs of edema and expansion of the compo-

nents of the lymphatic vessels are noted. The number of ves-

sels is decreased, but the vessels became larger and have scle-

rosed walls with perivascular leukocytic infiltration. Hematox-

ylin and eosin. Magnification × 40.

Figure 10. The fragment of Figure 9. The newly-formed blood

vessels with thin walls and red cells located in the connective

tissue on the place of reticular layer of dermis. In these vessels

the endothelial lining can be seen along the full length, the

remaining layers of the wall (muscle, adventitia) are absent.

Magnification × 400.

defect 6 weeks after the introduction of APCPB were

also found atrophy of the epidermis and hyperkeratosis,

but the severity of atrophy was significantly less. Also

skin appendages and all layers of the dermis were absent.

However, there was a formatting and some ordering of

collagen fibers in the connective tissue on the place of

dermis, there was significantly more vessels, reduced the

severity of edema and the degree of perivascular leuko-

cyte infiltration. In cytogram of such small infiltrates the

lymphocytes and macrophages dominated.

In 5 months after treatment, sclerosis of the dermis

was still decreased. The number of vessels decreased,

but the remaining vessels were large. Leukocytic infil-

trates around the vessels were present again and ap-

peared signs of edema.

Patient S., 30 years old (pregnancy 9 years ago): in

skin before the injection of APCPB were found hyper-

keratosis, atrophy of the papillary and reticular layers of

the dermis and smoothing, to almost complete disap-

pearance, of dermal papillae. Also hair follicles, sudori-

ferous and sebaceous glands were absent completely.

The dermis was replaced by scar connective tissue with

a chaotic arrangement of thick fibers and large cavities

with a transparent content and a small amount of blood

vessels, around which were noted perivascular edema

and leukocytic infiltration with a predominance of macro-

phages.

In the study of biopsy specimen from the edge of the

defect in 6 weeks after the injections of APCPB there

were found almost normal skin structure—epidermis,

papillary and reticular layers of the dermis. In this case,

only hyperkeratosis, small lymphocytic-macrophage infil-

tration of the vascular walls, a moderate perivascular and

tissue edema attracted attention. Also the number of

small vessels increased.

5. DISCUSSIONS

Thus, all patients had similar initial state o f skin striae

after pregnancy, (of course, the severity of the skin

changes was different) and was consistent with the lite-

rature data devoted to the description of “old” striae,

which are characterized by the thinning of the connec-

tive tissue stroma, which leads to the formation of lines

of atrophied skin, and the presence of a large number of

thickened elastin fibers [5,6,8,13].

Due to thinning and sclerosis of the dermis, which

remains over a long period of time, its blood supply is

disturbed. As a result of vascular disturbance there is no

proper nutrition and oxygenation of cells. There are

formed leukocytic infiltrates for lysis of nonviable tissue.

Scar changes gradually pr ogress, and all stru ctures of the

dermis, as the result of fibroblasts functional d isturbance

[5-10,13], with time are replaced by randomly arranged

thick fibers of collagen and elastin.

After the introduction of APCPB the edema of the

dermis and atrophy of the epithelium may become stronger

or weaker. However, after the procedure, in all observa-

tions in the connective tissue on the place of dermis

there is an increase of number of vessels, thinning and

ordering of distribution of collagen and elastin fibers.

I. Maiborodin et al. / Health 2 (2010) 1430-1436

1435

Apparently, injected cells somehow stimulate the

growth of blood vessels, perhaps participate in this

process because of the presence of pluripotent cells

which were already stimulated to differentiate into en-

dothelial or pericytal directions. After increasing the

number of vessels, especially the “young” with thin

walls, metabolic processes in the diseased tissues are

improve. As a result of optimizing cond itions of life and

functioning of the fibroblasts and tissue leukocytes the

metabolism of extracellular matrix components of the

connective tissue of the dermis intensifies, there is a re-

juvenation of collagen and elastin fibers, which is mani-

fested in the appearance of finer structures and ordering

their location [13]. Also , wh en metabolism impro v es and

therefore the volume of nonviable tissue reduces, perivas-

cular infiltration and its activity are decreased.

It should be noted that in the remote period of up to 5

months after the introduction of APCPB the number of

vessels somewhat is decreased, but their size still re-

mains bigger than original. The processes of neoangi-

ogenesis at this time does not stop and is quite active,

evidence of that can be the presence of wide blood ves-

sels with thin walls even at this period.

As a re sult of these therapeu tic procedur es some po si-

tive aesthetic results were obtained due to the fact that

stretch marks appear not as “sunken, hollow”, become

narrower. In addition, the state of the surrounding skin

improves, it becomes denser. Making this procedure, we

did not attempt to achiev e a significant therapeutic effect,

which manifests itself in full or partial disappearance of

skin defects. Use of cellular technology improved the

microcirculation in “old” stretch marks. The scars, ex-

isting for many years, according to the morphological

pattern following the introduction of APCPB, in particu-

lar, in number and size of vessels, in appearance fibers

of extracellular matrix of the dermis, are similar to

“young” scar.

After rejuvenation of the stretch marks it is possible to

perform some cosmetic procedures (mechanical, chemi-

cal or laser microdermabrasion (grinding) of the skin),

aimed at making these scars to less visible or even com-

pletely to eliminate them, as only fresh stretch marks,

which appeared less than a year ago, can be corrected

effectively [15,16].

Despite the fact that there are possibilities to struggle

against stretch marks such as laser grinding, and peeling

with the use of corrosive acids and microdermabrasion,

as well as their combinations, the prevention of stretch

marks is difficult process, but this is more effective than

treatment of already existing stretch marks [17-19].

6. CONCLUSIONS

Thereby, in abdominal skin striae after pregnancy,

completed several years ago, were found atrophy of the

epithelium, hyperkeratosis and sclerosis of all layers of

the dermis with smoothing of papillae, the decrease of

number of blood vessels and skin appendages. After the

introduction of APCPB the first visual signs of im-

provement of appearance of atrophic scars were detected

in 6-8 weeks. In 6 months there is well visible th e filling

of scars from the inside, the color of stretch marks be-

come closer to the color of normal skin, the “old”

atrophic scars begin to get sun tan. The depth of the

sunken tissue is reduced, scar skin becomes more dense,

less thinned on appearance. According to morphological

data after the application of APCPB the edema of the

dermis and atrophy of the epithelium may become

stronger or weaker. But in all cases in the connective

tissue on the place of dermis there is increasing of num-

ber of blood vessels due to the process of neoangiogene-

sis, reducing the degree of leukocytic infiltration, thin-

ning and ordering distribution of collagen and elastin

fibers, which enables more successful application of

cosmetic procedures for the correction of skin defects.

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