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Study on the effectiveness of sperm retrieval and relationship between a number of factors and sperm retrieval ability of microdissection testicular spem extraction technique on non

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Microdissection testicular spem extraction is currently the optimal sperm retrieval method for non-obstructive azoospermia patients. Research and development of this technique will help clinicians have more tools to collect sperm effectively for non-obstructive azoospermia patients, enabling these patients to have children of their own.

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STUDY ON THE EFFECTIVENESS OF SPERM RETRIEVAL AND

RELATIONSHIP BETWEEN A NUMBER OF FACTORS AND

SPERM RETRIEVAL ABILITY OF MICRODISSECTION

TESTICULAR SPEM EXTRACTION TECHNIQUE ON NON-OBSTRUCTIVE AZOOSPERMIA PATIENTS

Vu Thi Thu Trang 1 ; Quach Thi Yen 2 ; Nguyen Dinh Tao 3 ; Trinh The Son 3

SUMMARY

Objectives: Microdissection testicular spem extraction is currently the optimal sperm retrieval

method for non-obstructive azoospermia patients Research and development of this technique

will help clinicians have more tools to collect sperm effectively for non-obstructive azoospermia

patients, enabling these patients to have children of their own Subjects and methods: 100

patients with non-obstructive azoospermia who underwent micro TESE from August 2016 to

October 2018 at Military Institute of Clinical Embryology and Histology, Vietnam Military Medical

University Results: The average age of patients was 32.21 ± 4.55; the average duration of

infertility was 4.77 ± 3.37 years; 93% of patients were primary infertility; testicular volume 6.57 ±

2.77 mL; the average concentration of FSH, LH, and testosterone were 20.30 ± 12.63 mIU/mL,

10.83 ± 6.26 mIU/mL amd 4.24 ± 2.40 ng/mL, respectively 19 patients (19%) had gene

abnormalities in AZF region; Sertoli cell-only syndrome accounted for the highest proportion in

the histopathological subgroup (50%); the rate of sperm collection was 37% Surgical testicular

volume, endocrine concentration, AZF gene abnormalities and histopathological lesions related

to sperm collection possibility There was no short complications after surgery Conclusions:

Microdissection testicular spem extraction was a safe sperm retrieval method with a sperm

retrieval rate of 37% Surgical testicular volume, endocrine concentration, AZF gene

abnormalities and histopathological lesions related to sperm retrieval capacity

* Keywords: Non-obstructive azoospermia; Sperm retrieval; Micro-dissection testicular

sperm extraction.

INTRODUCTION

The appearence of intra cytoplasmic

sperm injection (ICSI) method in 1992 and

the success of the ICSI case with sperm

obtained from the epididymis aspiration of

azoospermia patients of Tournaye in 1994

has opened a revolution in the treatment

of male infertility especially with the azoospermia case Since then, there have been many different methods of retrieving sperm, each method has its own advantages and disadvantages,but for non-obstructive azoospermia patients,

2 Vietnam University of Traditional Medicine

3 Vietnam Military Medical University

Corresponding author: Vu Thi Thu Trang (drtrangvu@gmail.com)

Date received: 15/10/2019

Date accepted: 06/12/2019

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micro-dissection testicular sperm extraction

(micro TESE) is the best method for

sperm retrieval As the first to implement

this technique, Schlege N.P (1999) showed

that the ability to collect sperm was from

42 to 63% [2] This method reduces

damages, reduces the effect of testicular

function by micro-surgical microscopy that

helps to see the sperm ducts and avoid

blood vessels This is a new technique

applied in Vietnam, so the objectives of

the topic were: Determine the sperm

retrieval ability and explore some factors

related to the sperm retrieval ability of

micro TESE method in non-obstructive

azoospermia patients

SUBJECTS AND METHODS

1 Subjects

100 patients with non-obstructive

azoospermia were conducted micro TESE

at Military Institute of Clinical Embryology

and Histology, Vietnam Military Medical

University from August 2016 to October

2018

* Selection criteria: Patients without

sperm in the semen according to WHO

(2010) [3], not retrograde ejaculation;

underwent percutaneuos epididymal sperm

aspiration (PESA) or microsurgical epidymal

sperm aspiration (MESA - Microsurgical

Epidymal Sperm Aspiration) to eliminate

the obstructive azoospermia cases

* Exclusion criteria: Cases of secondary

hypogonadism; cases of acute diseases,

social diseases, endocrine diseases,

taking drugs and chemicals affecting

sperm production

2 Method Prospective, cohort study

RESULTS

1 Some characteristics of research subjects

* Age, infertility type, infertility period of the study subjects:

- The average age of the patient was 32.21 ± 4.55 The highest was 47 years old, the lowest was 21 years old Most in age group of > 30 - 40 years old (57%)

- Primary infertility accounted for 93% Only 7% were secondary infertility

- The average infertility period was 4.77 ± 3.37 years Patients with the longest infertility period were 19 years and the shortest were 1 year Infertility group from 2 - < 5 years was seen the most (52%), followed by group 5 - < 10 years (31%), the lowest was infertility group under 2 years (7%)

* The surgical testicular volume:

The average surgical testicular volume was 6.57 ± 2.77 mL The biggest testicular volume undergoing surgery was

16 mL, the smallest was 2 mL There were 59% testicles with volume from 5 -

< 10 mL; group with volume < 5 mL accounted for 27%; the lowest was the group with volume over 15 mL (1%)

* Concentrations of some hormones of study patients:

The average concentration of FSH, LH and testosterone of the study subjects were 20.30 ± 12.63 mIU/mL, 10.83 ± 6.26 mIU/mL and 4.24 ± 2.40 ng/mL

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* Results of AZF genetic testing:

Table 1:

Types of AZF gene abnormalities

The amount and

proportion/abnormality AZF gene

Abnormal AZF genes

19% of patients had AZF gene damage Associated lesions were the most common (42.1%), followed by AZFc gene abnormality (36.8%), the lowest was AZFa gene

abnormality (5.3%)

2 Results of retrieving sperm by micro TESE method in study patients

* Percentage of patients retrieving sperm:

Of the 100 patients participating in the study, 37 patients (37%) obtained sperm and

63 patients (63%) did not obtaine sperm

Table 2: Percentage of patients with AZF gene abnormality obtained sperm

Obtaining sperm (n = 7)

Not obtaining sperm (n = 12)

Abnormal AZF genes

(n = 19)

Among19 cases with abnormal AZF genes, abnormality in AZFc gene had the highest sperm retrieval rate (71.4%) The group with abnormality in AZF genes and

associated lesions were unable to find sperm

3 Relationship of some factors with the ability to collecting sperm from micro TESE

* Relationship between age, type of infertility, time/period of infertility, and ability of retrieving sperm:

The test results did not find the relationship between age, type of infertility, time of infertility and the ability of retrieving sperm of the micro TESE method in the study patient (p > 0.05)

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* Relationship between surgical testicular volume and the ability to retrieve sperm:

Table 3:

Obtaining sperm (n = 37)

Not obtaining sperm (n = 63) Surgical testicular volume

(mL)

p

OR (95%CI)

0.033 2.45 (1.06 - 5.62)

(*: χ2 test)

There was a relationship between testicular volume of ≥ 7 mL with surgery to see

sperm Subjects with testicular volume of ≥ 7 mL were more likely to find sperm 2.45 times higher than those with a testicular volume of less than 7 mL (p < 0.05)

* Relationship between several tests and the ability to obtain sperm:

Table 4: Relationship between FSH, LH, and testosterone concentrations and sperm

retrieval ability

Obtaining sperm (n = 37)

Not obtaining sperm (n = 63)

Hormone level

p

OR (95%CI)

FSH outside the normal value 24 (30.8) 54 (69.2)

0.015*

3.25 (1.22 - 8.63)

0.086**

Testosterone normal (2.2 - 8.0 ng/mL) 35 (43.8) 45 (56.2)

Testosterone outside the normal value 2 (10.0) 18 (90.0)

0.005**

(*: χ2 test; **: Fisher’s exact test)

There was a relationship between FSH concentration in the normal range and

testosterone level in the normal range with the surgery to see sperm (p < 0.05) We did

not find the relationship between normal LH concentration and the surgery to see

sperm (p > 0.05)

Table 5: Relationship between histopathological results and sperm retrieval ability

Histopathological results

Seminiferous tubule hyalinization 1 (9.1) 10 (90.9)

< 0.001**

χ2 = 20.31

(**: Fisher’s exact test)

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The group with impaired spermatogenesis

was most likely to obtain sperm at the

highest rate (70.4%), followed by the

halfway spermatogenesis group (41.7%)

and the lowest was the hyalinization

spermatomy tube group with the rate of

obtaining sperm of 9.1%

4 Accidents and complications

after surgery

There were no complications after surgery

such as: bleeding, hematoma, infection

DISCUSSION

Through the study results, we realized

that micro TESE method was an effective

method of collecting sperm for

non-obstructive azoospermia patients The

percentage of sperm obtained from

non-obstructive azoospermia patients by this

method reached 37% Micro TESE was

also a safe method, with small volume of

testicle tissue removed, each time only

from 5 to 10 mg of testicular tissue, thus

less affecting the testicular function after

surgery In the study, after the testicular

tissue sample was obtained under a

micro-surgical microscope, a sample of

collagenase type IA was added at a

concentration of 0.8 mg/mL, so that the

sperm cell line was separated from the

wall of the spermatogenesis tube This is

the first procedure implemented in

Vietnam instead of studying testicular

tissue with two prism or two obtuse

needles Dabaja A.A et al (2013)

recommended the use of enzymes to

process testicular tissue samples to

increase the chances of sperm retrieval

from tissue samples obtained [4]

However, compared to results published

by some authors in the world, our success rate was still lower Schlege N.P (1999) announced that the percentage of sperm collection by micro TESE method was about 42 - 63%; Bryon F.C (2014) showed that this rate reached 50 - 60% [2, 5] This difference may be due to patient selection criteria, the sample size, and it also depends on the surgical base and surgeon's experience

Compared with the sperm collection rate from testicles of non-obstructive azoospermia patients by TESE in Vietnam published by Trinh The Son et al (2015), the micro TESE method was the method

of collecting sperm from testicles much more effective than other methods (37% compared to 23.3%) [1] Thus, with the success of the micro-TESE, it has contributed to improving the quality and effectiveness of infertility treatment in general and male infertility in particular and providing opportunities for non-obstructive azoospermia patients to have their own baby

In the study, the AZFc gene abnormality

group had a higher sperm collection rate

than the other AZF gene abnormalities

that contributed to the counseling and prognosis for patients

The average FSH concentration of the study patients was 20.30 ± 12.63 mIU/m, much higher than the normal index of about 2 - 10 mIU/mL In this study, FSH and testosterone levels were correlated

with sperm retrieval ability (table 4) However,

some other authors, such as Kalsi J et al (2012) found that FSH concentrations did not have a prognostic value of sperm

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retrieval ability of micro TESE method [6];

Bermie M.A (2013), Bryson F.C (2014)

supposed that there was no single factor

that can assess the sperm retrieval ability

of micro TESE, so it is necessary to

combine all factors/elements to predict

[5, 7] This difference may be due to

differences in sample size and subjects

Among the histopathologic lesions, the

group with impaired spermatogenesis was

most likely to obtain sperm at the highest

rate (70.4%), followed by the halfway

spermatogenesis group (41.7%) and the

lowest was the hyalinization spermatomy

tube group with the rate of obtaining sperm

of 9.1% (table 5) Thus, histopathology

was a valuable factor in the prognosis of

sperm retrieval ability of micro TESE

CONCLUSION

Micro TESE is a method to collect

sperm from the testicle safely, effectively

in patients with non-obstructive azoospermia,

the sperm collection rate reacheed 37%

Testicular volumes, FSH concentration,

testosterone, AZF gene abnormalities,

histopathological lesions had a prognostic

value of sperm retrieval ability in

non-obstructive azoospermia patients by micro

TESE method

REFERENCES

1 Trinh The Son, Vu Van Tam. Evaluation

of results of percutaneuos epididymal sperm aspiration (PESA) and testicular spem extraction (TESE) on azoospermia patients in Haiphong Obstetrics and Gynecology Hospital Journal of Military Medicine 2015

2 Schlegel P.N Testicular sperm extraction: Micro-dissection improves sperm yield with minimal tissue excision Hum Reprod 1999,

14, (1), pp.131-135

3 WHO. WHO laboratory manual for the examination and processing of human semen Fifth edition Switzerland 2010

4 Dabaja A.A, Schlegel N.P Micro-dissection testicular sperm extraction: An update Asian Jourmal of Andrology 2013, 15, pp.35-39

5 Bryson F.C, Ramasamy R, Sheehan M, Palermo D.G, Rosenwaks Z, Schlegel N.P

Severe testicular atrophy does not affect the success of micro-dessection testicular sperm extraction J Urol 2014, 191 (1), pp.175-178

6 Kalsi J, Thum Y.M, Muneer A, Abdullah

H, Minhas S. In the era of micro-dissection sperm retrieval (m-TESE) is an isolated testicular biopsy necessary in the management

of men with non-obstructive azoospermia BJU 2012, 109 (3), pp 418-424

7 Bernie M.A, Ramasamy R, Schlegel

micro-dissection testicular sperm extraction Clinical Andrology 2013, 23, p.5

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